Methods of using il-1 antagonists to treat alzheimer&#39;s disease

ABSTRACT

The invention provides methods of treating, inhibiting, or ameliorating a disease characterized by aberrant deposition of beta amyloid in a subject in need thereof, comprising administering to a subject a therapeutic amount of an interleukin 1 (IL-1) antagonist, wherein the disease, or condition is treated, inhibited, or ameliorated, or wherein the onset or progression of the disease, or at least one symptom of the disease, is delayed. The IL-1 antagonist is an IL-1 trap, preferably comprising a sequence selected from the group consisting of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, and 28 and capable of binding and inhibiting IL-1. The therapeutic methods are useful for treating a human adult suffering from Alzheimer&#39;s Disease or cerebral amyloid angiopathy.

FIELD OF THE INVENTION

The invention relates to methods of using an interleukin-1 (IL-1)antagonist to treat or to slow the progression of a diseasecharacterized in part by beta amyloid (Aβ) expression or activity, or byaberrant deposition of beta amyloid in a subject, such as in Alzheimer'sdisease, and more specifically, the pathologies associated with such adisease, including for example, behavioral changes or cognitivedysfunction associated with Alzheimer's disease.

STATEMENT OF RELATED ART

The proinflammatory cytokine interleukin-1 (IL-1) is an important playerin inflammatory processes throughout the body, including in the centralnervous system (CNS). IL-1 is found in two distinct isoforms, IL-1a andIL-β, although IL-β is considered the primary active isoform.Upregulation of IL-β is part of the response to a range of CNS insults,including infections, stroke, and traumatic injuries (Allan, S. M.,Tyrrell, P. J., & Rothwell, N. J. (2005), Nature Reviews Immunology, 5,629-640). This neuroinflammatory response is characterized by activationof resident glial cells (microglia and astrocytes), infiltration ofperipheral immune cells, and the expression of inflammatory mediators,such as cytokines and chemokines (Shaftel, S. S., Griffin, W. S. T., &O'Banion, M. K. (2008), Journal of Neuroinflammation, 5:7).

IL-1-mediated neuroinflammation may also play a role in the pathogenesisof neurodegenerative diseases. For example, elevated levels of IL-1 arereported in the brain tissue of patients with Alzheimer's disease (AD).AD (Griffin, W. S., Stanley, L. C., Ling, C., White, L., MacLeod, V.,Perrot, L. J., Araoz, C. (1989), Proceedings of the National Academy ofSciences of the United States of America, 86, 7611-7615) and in rodentmodels of the disease (Benzing, W. C., Wujek, J. R., Ward, E. K.,Shaffer, D., Ashe, K. H., Younkin, S. G., & Brunden, K. R. (1999),Neurobiology of Aging, 20(6), 581-589). The degree of neuroinflammationand IL-1 expression has been shown to correlate with the level ofpathology in AD patients (Sheng, J. G., Ito, K., Skinner, R. D., Mrak,R. E., Rovnaghi, C. R., Van Eldik, L. J., & Griffin, W. S. (1996),Neurobiology of Aging, 17(5), 761-766).

Although the research to date may support a link between AD and IL-1, itis unclear what role IL-1 may play. IL-1 modulates actions thatcontribute to AD pathology, including the synthesis and processing ofthe Aβ precursor protein and the activity of acetylcholinesterase (Mrak,R. E. & Griffin, W. S. (2001), Neurobiology of Aging, 22(6), 903-908).Chronic IL-1 expression has been associated with demyelination (Ferrari,C. C., Depino, A. M., Prada, F., Muraro, N., Camptbell, S., Podhajcer,O., Pitossi, F. J. (2004), American Journal of Pathology, 165(5),1827-1837), breakdown of the blood-brain barrier, and neutrophilrecruitment (Ferrari et al, supra; Shaftel, S. S., Carlson, T. J.,Olschowka, J. A., Kyrkanides, S., Matousek, S. B., & O'Banion, M. K.(2007), Journal of Neuroscience, 27(35), 9301-9309). IL-1 also activatesmicroglia, which in turn produce pro-inflammatory cytokines such asIL-1, IL-6, and tumor necrosis factor alpha (TNFα). Neuronal insults,such as the accumulation of Aβ, may therefore induce a self-propagatingcycle of cytokine activation in which levels of IL-1 constantly rise,leading to neuronal damage and further plaque deposition (Griffin, W. S.T., Sheng, J. G., Royston, M. C., Gentelman, S. M., McKenzie, J. E.,Graham, D. I., Mrak, R. E. (1998), Brain Pathology, 8, 65-72).

Prior research has explored the use of anti-inflammatory drugs as atherapeutic strategy against AD. Epidemiological studies havedemonstrated a reduced risk of developing the disease in long-termnon-steroidal anti-inflammatory drug (NSAID) users (Szekely, C. A.,Breitner, J. C. S., Fitzpatrick, A. L., Rea, T. D., Psaty, B. M.,Kuller, L. H., & Zandi, P. P. (2008), Neurology, 70(1), 17-24). Intransgenic animal models, chronic NSAID administration has been somewhateffective in preventing or delaying the onset of amyloid deposition,dystrophic neurite formation and inflammation (Lim, G. P., Yang, F.,Chu, T., Chen, P., Beech, W., Teter, B., Cole, G. M. (2000), Journal ofNeuroscience, 20(15), 5709-5714). However, randomized clinical trialshave failed to consistently support the therapeutic effectiveness ofNSAIDs against AD (Scharf, S., Mander, A., Ugoni, A., Vajda, F., &Christophidis, N. (1999), Neurology, 53(1), 197-201; Aisen, P. S.,Schafer, K. A., Grundman, M., Pfeiffer, E., Sano, M., Davis, K. L.,Thal, L. J. (2003), Journal of the American Medical Association,289(21), 2819-2826).

Given the small number of approved therapies to treat, or to slow downthe progression of a disease characterized in part by beta amyloidexpression, activity, or aberrant deposition, such as AD, there is aneed to identify and explore the use of other agents for treating thesediseases, such as the IL-1 antagonists as described herein.

BRIEF SUMMARY OF THE INVENTION

The invention provides a method for treating a subject suffering from adisease characterized in part by the deposition and/or activity of betaamyloid in the brain tissue of a subject by administering aninterleukin-1 (IL-1) antagonist. An IL-1 antagonist is a compoundcapable of blocking or inhibiting at least one biological activity ofIL-1. An IL-1 antagonist may take the form of an antibody, a solublereceptor, or a fusion protein capable of trapping IL-1, such as an IL-1trap as described herein. In one embodiment, the subject is a humanpatient suffering from Alzheimer's Disease (AD). The IL-1 trap may beadministered alone, or in conjunction with one or more therapeuticagents that are useful for treating AD, or for slowing the progressionof the disease, or for ameliorating at least one symptom associated withthe disease, including, but not limited to behavioral changes associatedwith AD, or the cognitive decline or dysfunction observed in patientswith AD.

Accordingly, in a first aspect, the invention features a method fortreating, or delaying the onset, or the progression of a diseasecharacterized in part by beta amyloid expression, activity, ordeposition in a subject in need thereof, or for ameliorating at leastone symptom associated with the disease, the method comprisingadministering to the subject a therapeutically effective amount of anIL-1 antagonist as a first therapeutic agent, wherein the IL-1antagonist is selected from the group consisting of an antibody specificfor IL-1 alpha or IL-1 beta, or an antigen-binding fragment thereof, asoluble IL-1 receptor, and an IL-1 trap, wherein the IL-1 trap is afusion protein comprising an IL-1 binding portion of the extracellulardomain of IL-1 RAcP, an IL-1 binding portion of the extracellular domainof IL-1 R1, and a multimerizing component.

In one embodiment, the invention provides a method for treating, ordelaying the onset, or the progression of a disease characterized inpart by beta amyloid expression, activity, or deposition in a subject inneed thereof, or for ameliorating at least one symptom associated withthe disease, the method comprising administering to the subject atherapeutically effective amount of an IL-1 antagonist as a firsttherapeutic agent, wherein the IL-1 antagonist is an IL-1 trap, whereinthe IL-1 trap is a fusion protein comprising an IL-1 binding portion ofthe extracellular domain of IL-1 RAcP, an IL-1 binding portion of theextracellular domain of IL-1 R1, and a multimerizing component.

In a related aspect, the invention features a method of inhibiting IL-1activity for treating a disease, or delaying the onset or theprogression of a disease characterized in part by beta amyloidexpression, activity, or deposition in a subject in need thereof, or forameliorating at least one symptom associated with the disease, themethod comprising administering to the subject a therapeuticallyeffective amount of an IL-1 antagonist as a first therapeutic agent.

In one embodiment, the first therapeutic agent is an IL-1 antagonistselected from the group consisting of an antibody specific for IL-1alpha or IL-1 beta, a soluble IL-1 receptor that blocks or inhibits theactivity of IL-1 alpha and/or beta, or an IL-1 fusion protein (e.g. anIL-1 trap as described herein).

In one particular embodiment, the IL-1 antagonist is a fusion proteincomprising an IL-1 binding portion of the extracellular domain of IL-1Receptor Accessory protein (IL-1RAcP), an IL-1 binding portion of theextracellular domain of IL-1 R1, and a multimerizing component.

In one embodiment, the IL-1 antagonist is an IL-1-specific fusionprotein comprising two IL-1 receptor components and a multimerizingcomponent, for example, an IL-1 trap as described in U.S. Pat. Nos.6,927,044; 6,472,179; 7,459,426; 8,414,876; 7,361,350; 8,114,394;7,820,154 and 7,632,490, all of which are specifically incorporated byreference in their entirety.

In one embodiment, the IL-1 trap is the fusion protein shown in SEQ IDNO: 2, 4, 6, 8, 10, 12,14, 16, 18, 20, 22, 24, 26 and 28. In oneembodiment, the IL-1 trap is shown in SEQ ID NO: 28. In one embodiment,the IL-1 trap is shown in SEQ ID NO: 10. The invention encompasses theuse of an IL-1 trap substantially identical to the protein of SEQ ID NO:2, 4, 6, 8, 10, 12,14, 16, 18, 20, 22, 24, 26, 28, that is, a proteinhaving at least 95% identity, at least 97% identity, at least 98%identity to the protein of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18,20, 22, 24, 26, and 28 and capable of binding and inhibiting IL-1.Further, in specific embodiments, the IL-1 antagonist is a modified IL-1trap comprising one or more receptor components and one or moreimmunoglobulin-derived components specific for IL-1 and/or an IL-1receptor. In another embodiment, the IL-1 antagonist is a modified IL-1trap comprising one or more immunoglobulin-derived components specificfor IL-1 and/or an IL-1 receptor.

The subject being treated is most preferably a human suffering from adisease associated with beta-amyloid deposition and/or activity in thebrain, such as Alzheimer's disease. Other subjects that may benefit fromsuch therapy include subjects suffering from multi-infarct dementia,cognitive impairment, Down's syndrome and cerebral amyloid angiopathy.In certain embodiments the Alzheimer's disease may be prodromal,preclinical or clinical stage AD. In certain embodiments the cerebralamyloid angiopathy may be preclinical or clinical stage cerebral amyloidangiopathy.

In one embodiment, the IL-1 trap is a fusion protein comprising theamino acid sequence of SEQ ID NO: 10.

In one embodiment, the IL-1 trap is a fusion protein comprising theamino acid sequence of SEQ ID NO: 28.

In certain embodiments, the administration of the IL-1 trap issubcutaneous, intramuscular, intranasal, intraarterial, intravenous,intrathecal, intraventricular, intracerebral, topical, transdermaladministration or oral.

In one embodiment, a therapeutically effective amount of the IL-1 trapto be administered is between about 1 mg/kg to about 750 mg/kg.

In one embodiment, a therapeutically effective amount of the IL-1 trapto be administered is between about 10 mg/kg to about 500 mg/kg.

In one embodiment, a therapeutically effective amount of the IL-1 trapto be administered is between about 50 mg/kg to about 150 mg/kg.

In certain embodiments the methods of the invention provide fortreating, inhibiting, or ameliorating a disease, or delaying the onset,or the progression of a disease characterized in part by beta amyloidexpression, activity, or deposition in a subject in need thereof byadministering a therapeutically effective amount of an IL-1antagonist/trap, as described herein, as a first therapeutic agent and atherapeutically effective amount of one or more other therapeuticagents, wherein the disease or at least one symptom associated with thedisease is lessened in severity or duration, or wherein the onset orprogression of the disease or at least one symptom associated with thedisease is delayed.

In certain embodiments the at least one symptom associated with thedisease is selected from the group consisting of memory loss,depression, anxiety, dementia, irritability, confusion, inattention,mood swings, and aggressive and/or apathetic behavior.

In certain embodiments the other therapeutic agent(s) is/areadministered by any route selected from subcutaneous, intramuscular,intranasal, intraarterial, intravenous, intrathecal, intraventricular,intracerebral, topical, transdermal administration or oral.

In one embodiment, the other therapeutic agent is anacetylcholinesterase inhibitor or a glutamate pathway modifier.

In one embodiment, the acetylcholinesterase inhibitor is selected fromthe group consisting of ARICEPT® (donepezil HCl), EXELON® (rivastigminetartrate), and RAZADYNE® (galantamine HBr).

In one embodiment, the glutamate pathway modifier is Namenda(memantine).

In one embodiment, the other therapeutic agent(s) is/are selected fromthe group consisting of a different IL-1 antagonist, ananti-inflammatory agent, an antibody specific for tau, an antibodyspecific for beta amyloid and a microtubule stabilizer.

In one embodiment, the other therapeutic agent(s) is/are a differentIL-1 antagonist selected from the group consisting of an IL-1 alpha orIL-1 beta antibody, a soluble IL-1 receptor, a different IL-1 trap,anakinra (KINERET®) and canakinumab.

In one embodiment, the anti-inflammatory agent is aspirin or a differentNSAID.

In one embodiment, the antibody specific for beta amyloid is selectedfrom the group consisting of solanezumab, gantenerumab, andbapineuzumab.

In one embodiment, the microtubule stabilizer is epothilone.

A second aspect provides a method of improving cognitive impairment in amammal having beta amyloid deposits in brain tissue, the methodcomprising administering to the subject a therapeutically effectiveamount of an IL-1 antagonist as a first therapeutic agent, wherein theIL-1 antagonist is selected from the group consisting of an antibodyspecific for IL-1 alpha or IL-1 beta, or an antigen binding fragmentthereof, a soluble IL-1 receptor, and an IL-1 fusion protein (IL-1trap).

In a related aspect, the invention provides a method of improvingcognitive impairment in a mammal having beta amyloid deposits in braintissue, the method comprising administering to the subject atherapeutically effective amount of an IL-1 antagonist as a firsttherapeutic agent, wherein the IL-1 antagonist is an IL-1 fusion protein(IL-1 trap).

In one embodiment, the IL-1 antagonist is a fusion protein comprising anIL-1 binding portion of the extracellular domain of IL-1 RAcP, an IL-1binding portion of the extracellular domain of IL-1 R1, and amultimerizing component, wherein the mammal demonstrates an improvementin cognitive function(s) without the necessity of a change in the betaamyloid plaque burden in the brain.

In one embodiment, the invention provides for a method of improvingcognitive impairment in a mammal having beta amyloid deposits in braintissue, the method comprising administering a composition comprising anIL-1 trap of the invention as a first therapeutic agent, either alone,or in combination with one or more other therapeutic agents useful fortreating the disease or at least one symptom of the disease. In oneembodiment, the method provides for improvement of cognitive impairmentin a subject having beta amyloid deposits in the brain, withoutnecessarily altering the amount (increase or decrease) of beta amyloidin the brain. The improvement of cognitive impairment in a subject maybe an improvement in learning performance, or an improvement in memoryperformance, or a decrease in memory loss, or a decrease in learningimpairment.

In one embodiment, the cognitive impairment is associated withAlzheimer's disease. In certain embodiments, the treatment results inslowing the progression of any one or more cognitive or non-cognitivebehavioral changes in the subject, including but not limited to memoryloss, inability to learn, depression, anxiety, dementia, irritability,confusion, inattention, mood swings, diminished general locomotor and/orexploratory activity and aggressive and/or apathetic behavior. Othersubjects that may benefit from therapy with an IL-1 trap of theinvention in combination with one or more other therapeutic agentsinclude subjects suffering from multi-infarct dementia, cognitiveimpairment, Down's syndrome and cerebral amyloid angiopathy. In certainembodiments the Alzheimer's disease may be prodromal, preclinical orclinical stage AD. In certain embodiments the cerebral amyloidangiopathy may be preclinical or clinical stage cerebral amyloidangiopathy.

In one embodiment, the IL-1 trap is the fusion protein shown in SEQ IDNO: 2, 4, 6, 8, 10, 12,14, 16, 18, 20, 22, 24, 26 and 28. In oneembodiment, the IL-1 trap is shown in SEQ ID NO: 28. In one embodiment,the IL-1 trap is shown in SEQ ID NO: 10. The invention encompasses theuse of an IL-1 trap substantially identical to the protein of SEQ ID NO:2, 4, 6, 8, 10, 12,14, 16, 18, 20, 22, 24, 26, 28, that is, a proteinhaving at least 95% identity, at least 97% identity, at least 98%identity to the protein of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18,20, 22, 24, 26, and 28 and capable of binding and inhibiting IL-1.Further, in specific embodiments, the IL-1 antagonist is a modified IL-1trap comprising one or more receptor components and one or moreimmunoglobulin-derived components specific for IL-1 and/or an IL-1receptor. In another embodiment, the IL-1 antagonist is a modified IL-1trap comprising one or more immunoglobulin-derived components specificfor IL-1 and/or an IL-1 receptor.

In certain embodiments, subjects being treated may suffer from chronicneuroinflammation, which may contribute to the neurodegeneration and/orassociated cognitive or non-cognitive dysfunction observed in patientswith Alzheimer's disease, or any of the other neurodegenerativeconditions described herein. In certain embodiments, the subjects beingtreated with the IL-1 trap of the invention may have improved cognitiveor non-cognitive behavioral symptoms following treatment, but willexhibit no change in the amount of beta-amyloid deposited in the brain.In certain embodiments, the subjects being treated with the IL-1 trap ofthe invention will demonstrate a diminished immune response triggered bythe amyloid plaque burden. The reduction in immune response may be shownby a reduction in the number, activated phenotype and/or the size ofpen-plaque microglia.

The methods of the invention include administration of the IL-1antagonist (IL-1 trap) by any means known to the art, for example,subcutaneous, intramuscular, intranasal, intraarterial, intravenous,intracerebral, intraventricular, intrathecal, topical, transvaginal,transdermal, transanal administration or oral routes of administration.

In one embodiment, a therapeutically effective amount of the IL-1antagonist (IL-1 trap) to be administered to a subject in need thereofranges from about 1 mg/kg to about 750 mg/kg, or about 10 mg/kg to about500 mg/kg, or more preferably from about 50 mg/kg to about 150 mg/kg. Inone embodiment, the IL-1 trap is administered on a weekly basis.

In one embodiment, a therapeutically effective amount of the IL-1antagonist (IL-1 trap) to be administered to a subject in need thereofranges from about 10 mg to about 500 mg, or about 100 mg to about 320mg. In one embodiment, a therapeutically effective amount of the IL-1antagonist (IL-1 trap) to be administered to a subject in need thereofis about 100 mg, or about 160 mg, or about 320 mg. In certainembodiments the IL-1 trap is administered on a weekly basis.

In certain embodiments of the therapeutic methods of the invention, thesubject is treated with a combination of an IL-1 trap and one or moreother (second or third, etc.) therapeutic agents. The other therapeuticagents may be a second IL-1 antagonist, such as, for example, anakinra(KINERET®) or canakinumab, or a second different IL-1 trap, or arecombinant, nonglycosylated form of the human IL-1 receptor antagonist(IL-1Ra), or an anti-IL-18 drug such as IL-18BP or a derivative, anIL-18 Trap, anti-IL-18, anti-IL-18R1, or anti-IL-18Racp. Otherco-therapies may include an acetylcholinesterase inhibitor (e.g.ARICEPT® (donepezil HCl), EXELON® (rivastigmine tartrate), RAZADYNE®(galantamine HBr)), or a glutamate pathway modifier, such as, Namenda(memantine HCl). Other co-therapies include aspirin or other NSAIDs, orother inflammatory inhibitors such as inhibitors of caspase-1, p38,IKK1/2, CTLA-4Ig, anti-IL-6 or anti-IL6Ra, etc. Other co-therapiesinclude an antibody specific for tau or an antibody specific for betaamyloid (such as solanezumab, gantenerumab, or bapineuzumab), as well asa microtubule stabilizer (such as epothilone B).

In a third aspect, the invention features a therapeutic method oftreating a disease characterized by deposition of beta-amyloid in asubject, or ameliorating at least one symptom of a disease characterizedby aberrant deposition of beta-amyloid in a subject, such as Alzheimer'sdisease, by administering a pharmaceutical composition comprising anIL-1 trap and a pharmaceutically acceptable carrier, in a dose range ofabout 1 mg/kg to about 300 mg/kg, preferably about 50 mg/kg to about 150mg/kg alone, or in combination with a second therapeutic agent usefulfor treating the disease.

In one embodiment, the invention provides for delaying the onset of, orslowing the progression of the disease, comprising administering to asubject in need thereof a pharmaceutical composition comprising an IL-1antagonist and a pharmaceutically acceptable carrier, in a dose range ofabout 10 mg/kg to about 300 mg/kg, or about 50 mg/kg to about 150 mg/kgon a weekly basis for a treatment period of between 1 week, 1 month, toone year or more. In certain embodiments, the treatment could last fordecades.

In certain embodiments, the IL-1 antagonist to be used as a first orsecond therapeutic agent is an antibody specific for either IL-1 alphaor IL-1 beta.

In certain embodiments the IL-1 antagonist to be used as a first orsecond therapeutic agent is a soluble IL-1 receptor that blocks orinhibits the activity of either or both IL-1 alpha and/or IL-1 beta.

In certain embodiments, the IL-1 antagonist to be used as a first orsecond therapeutic agent is anakinra or canakinumab.

In certain embodiments, the IL-1 antagonist to be used as a first orsecond therapeutic agent is an IL-1 trap as described herein.

In one particular embodiment, the IL-1 antagonist to be used as a firstor second therapeutic agent is the fusion protein (IL-1 trap) as shownin SEQ ID NO: 2, 4, 6, 8, 10, 12,14, 16, 18, 20, 22, 24, 26 or 28. Inone embodiment, the IL-1 trap is shown in SEQ ID NO: 28. In oneembodiment, the IL-1 trap is shown in SEQ ID NO: 10. The inventionencompasses the use of an IL-1 trap substantially identical to theprotein of SEQ ID NO: 2, 4, 6, 8, 10, 12,14, 16, 18, 20, 22, 24, 26, 28,that is, a protein having at least 95% identity, at least 97% identity,at least 98% identity to the protein of SEQ ID NO: 2, 4, 6, 8, 10, 12,14, 16, 18, 20, 22, 24, 26, and 28 and capable of binding and inhibitingIL-1. Further, in specific embodiments, the IL-1 antagonist is amodified IL-1 trap comprising one or more receptor components and one ormore immunoglobulin-derived components specific for IL-1 and/or an IL-1receptor. In another embodiment, the IL-1 antagonist is a modified IL-1trap comprising one or more immunoglobulin-derived components specificfor IL-1 and/or an IL-1 receptor.

In certain embodiments, the IL-1 antagonist may be administered twice aweek, or weekly, or monthly, or bi-monthly, or less frequently dependingon the results achieved.

In certain embodiments, the doses may be adjusted if it is determinedthat the patient may need chronic life-long therapy with the IL-1 trapalone, or in conjunction with a second therapeutic agent useful fortreating the disease. In certain embodiments, the methods for treating adisease characterized in part by beta amyloid activity or deposition inbrain tissue of a patient comprises administering to a subject in needthereof a pharmaceutical composition comprising an IL-1 trap at doses ofabout 100 mg, or about 160 mg, or about 320 mg and a pharmaceuticallyacceptable carrier. In certain embodiments the IL-1 trap is administeredon a weekly basis. In certain embodiments, the IL-1 trap may beadministered on a bi-weekly basis, a monthly basis, or a bi-monthlybasis, or less frequently as determined by a patient's response totherapy.

In certain embodiments, the pharmaceutical composition may contain asecond or third therapeutically effective amount of another agent usefulfor treating the disease (a co-formulation). The second or third othertherapeutic agent(s) may be a second IL-1 antagonist, such as, forexample, anakinra (KINERET®) or canakinumab, a different IL-1 trap, arecombinant, nonglycosylated form of the human IL-1 receptor antagonist(IL-1Ra), or an anti-IL-18 drug such as IL-18BP or a derivative, anIL-18 Trap, anti-IL-18, anti-IL-18R1, or anti-IL-18Racp. Otherco-therapies include acetylcholinesterase inhibitors (e.g. ARICEPT®(donepezil HCl), EXELON® (rivastigmine tartrate), RAZADYNE® (galantamineHBr)), or glutamate pathway modifiers, such as, Namenda (memantine HCl).Other co-therapies include aspirin or other NSAIDs, or otherinflammatory inhibitors such as inhibitors of caspase-1, p38, IKK1/2,CTLA-4lg, anti-IL-6 or anti-IL6Ra, etc. Other co-therapies include anantibody specific for tau or an antibody specific for beta amyloid (suchas solanezumab, gantenerumab, or bapineuzumab), as well as a microtubulestabilizer (such as epothilone B).

In one embodiment, the first and second other therapeutic agent may beadministered simultaneously in one pharmaceutical formulation, or may beadministered sequentially in different pharmaceutical compositions.

In one embodiment, the disease that is to be treated with apharmaceutical composition containing an IL-1 trap of the invention isAlzheimer's disease. In certain embodiments, the treatment with thepharmaceutical composition results in preventing the onset of, slowingthe progression of, or ameliorating/improving any one or more cognitiveor non-cognitive behavioral changes in the subject suffering fromAlzheimer's disease, including but not limited to memory loss, inabilityto learn, depression, anxiety, dementia, inattention, irritability,confusion, mood swings and aggressive and/or apathetic behavior.

A further aspect of the invention provides for the use of an IL-1antagonist of the invention for treating, or delaying the onset, or theprogression of a disease characterized in part by beta amyloidexpression, activity, or deposition in a subject in need thereof, or forameliorating at least one symptom associated with the disease, themethod comprising administering to the subject a therapeuticallyeffective amount of an IL-1 antagonist as a first therapeutic agent. Incertain embodiments, the IL-1 antagonist may be an antibody or solublereceptor that inhibits the activity of either IL-1 alpha or IL-1 beta,or it may be an IL-1 trap as described herein having the sequences asset forth in SEQ ID NOs: 2, 4, 6, 8, 10, 12,14, 16, 18, 20, 22, 24, 26or 28. The IL-1 antagonist may be used alone or in conjunction with oneor more other therapeutic agents that block, inhibit or antagonizeeither or both IL-1 alpha or IL-1 beta.

In one embodiment, the IL-1 antagonist is a fusion protein comprising anIL-1 binding portion of the extracellular domain of IL-1 RAcP, an IL-1binding portion of the extracellular domain of IL-1 R1, and amultimerizing component.

In a related aspect, the invention provides for the use of an IL-1antagonist of the invention for the preparation of a medicament fortreating, or delaying the onset, or the progression of a diseasecharacterized in part by beta amyloid expression, activity, ordeposition in a subject in need thereof, or for ameliorating at leastone symptom associated with the disease, the method comprisingadministering to the subject a therapeutically effective amount of anIL-1 antagonist as a first therapeutic agent. In certain embodiments,the IL-1 antagonist may be an antibody specific for IL-1 alpha or IL-1beta, or it may be a soluble receptor that blocks or inhibits theactivity of either IL-1 alpha and/or IL-1 beta, or it may be a fusionprotein comprising an IL-1 binding portion of the extracellular domainof IL-1 RAcP, an IL-1 binding portion of the extracellular domain ofIL-1 R1, and a multimerizing component.

In one embodiment, the IL-1 fusion protein comprising an IL-1 bindingportion of the extracellular domain of IL-1 RAcP, an IL-1 bindingportion of the extracellular domain of IL-1 R1, and a multimerizingcomponent is the IL-1 trap shown in any of SEQ ID NOs: 2, 4, 6, 8, 10,12,14, 16, 18, 20, 22, 24, 26 and 28. In one embodiment, the IL-1 trapis shown in SEQ ID NO: 28. In one embodiment, the IL-1 trap is shown inSEQ ID NO: 10. The invention encompasses the use of an IL-1 trapsubstantially identical to the protein of SEQ ID NO: 2, 4, 6, 8, 10,12,14, 16, 18, 20, 22, 24, 26, 28, that is, a protein having at least95% identity, at least 97% identity, at least 98% identity to theprotein of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26,and 28 and capable of binding and inhibiting IL-1. Further, in specificembodiments, the IL-1 antagonist is a modified IL-1 trap comprising oneor more receptor components and one or more immunoglobulin-derivedcomponents specific for IL-1 and/or an IL-1 receptor. In anotherembodiment, the IL-1 antagonist is a modified IL-1 trap comprising oneor more immunoglobulin-derived components specific for IL-1 and/or anIL-1 receptor.

Other objects and advantages will become apparent from a review of theensuing detailed description.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1a shows the results of the water maze acquisition test.

FIG. 1b shows the results of the water maze retention test (Time in GoalQuadrant).

FIG. 1c shows the results of the water maze retention test (Platformcrosses).

FIG. 2 shows the results of the open field test.

FIG. 3 shows increased plaque burden in transgenic animals.

FIG. 4a shows the median number of microglia-like cells per plaque.

FIG. 4b shows the mean size of lba-1-immunoreactive microglial-likecells in transgenic mice stratified by proximity to plaques.

FIG. 4c shows the mean difference in microglial size per animal inmicroglial in normal tissue (at least 30 microns from the nearestplaque) versus contacting amyloid plaques.

FIG. 5 shows the mean dorsal hippocampal volume at sacrifice for thefour groups of animals.

FIG. 6a shows the nucleic acid sequence (SEQ ID NO: 27) of the mouseIL-1 trap and FIG. 6b shows the amino acid sequence (SEQ ID NO: 28) ofthe mouse IL-1 trap as utilized in the studies described herein.

DETAILED DESCRIPTION

Before the present methods are described, it is to be understood thatthis invention is not limited to particular methods, and experimentalconditions described, as such methods and conditions may vary. It isalso to be understood that the terminology used herein is for thepurpose of describing particular embodiments only, and is not intendedto be limiting, since the scope of the present invention will be limitedonly the appended claims.

As used in this specification and the appended claims, the singularforms “a”, “an”, and “the” include plural references unless the contextclearly dictates otherwise. Thus for example, a reference to “a method”includes one or more methods, and/or steps of the type described hereinand/or which will become apparent to those persons skilled in the artupon reading this disclosure and so forth.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of the present invention, the preferred methodsand materials are now described. All patents, applications andnon-patent publications mentioned in this specification are incorporatedherein by reference in their entireties.

General Description

Alzheimer's disease (AD) is a degenerative brain disorder characterizedclinically by both cognitive and non-cognitive behavioral changes,including progressive memory deficits, depression, anxiety, dementia,irritability, mood swings, inattention, aggressive and/or apatheticbehavior, confusion, gradual physical deterioration and, ultimately,death. Histologically, the disease is characterized by neuritic plaques,composed primarily of beta amyloid (Aβ) peptide. The plaques are foundprimarily in the association cortex, limbic system and basal ganglia.Beta amyloid peptide is the cleavage product of beta amyloid precursorprotein (β APP or APP). APP is a type I transmembrane glycoprotein thatcontains a large ectopic N-terminal domain, a transmembrane domain, anda small cytoplasmic C-terminal tail. Alternative splicing of thetranscript of the single APP gene on chromosome 21 results in severalisoforms that differ in the number of amino acids.

It is believed that Aβ may play a role in the neuropathology ofAlzheimer's disease. For example, familial forms of the disease havebeen linked to mutations in APP and the presenilin genes (Tanzi et al.,1996, Neurobiol. Dis. 3:159-168; Hardy, 1996, Ann. Med. 28:255-258).Furthermore, diseased-linked mutations in these genes result inincreased production of the 42-amino acid form of Aβ, the predominantform found in amyloid plaques.

The proinflammatory cytokine interleukin-1 (IL-1) is as an importantplayer in inflammatory processes throughout the body, including in thecentral nervous system (CNS). IL-1 is found in two distinct isoforms,IL-1α and IL-1β, although IL-1β is considered the primary activeisoform. Upregulation of IL-1β is part of the response to a range of CNSinsults, including infections, stroke, and traumatic injuries (Allan, S.M., Tyrrell, P. J., & Rothwell, N. J. (2005). Nature Reviews Immunology,5, 629-640). This neuroinflammatory response is characterized byactivation of resident glial cells (microglia and astrocytes),infiltration of peripheral immune cells, and the expression ofinflammatory mediators, such as cytokines and chemokines (Shaftel, S.S., Griffin, W. S. T., & O'Banion, M. K. (2008), Journal ofNeuroinflammation, 5:7).

IL-1-mediated neuroinflammation may also play a role in the pathogenesisof neurodegenerative diseases (Griffin, W. S., Stanley, L. C., Ling, C.,White, L., MacLeod, V., Perrot, L. J., Araoz, C. (1989), Proceedings ofthe National Academy of Sciences of the United States of America, 86,7611-7615; Benzing, W. C., Wujek, J. R., Ward, E. K., Shaffer, D., Ashe,K. H., Younkin, S. G., & Brunden, K. R. (1999), Neurobiology of Aging,20(6), 581-589; Sheng, J. G., Ito, K., Skinner, R. D., Mrak, R. E.,Rovnaghi, C. R., Van Eldik, L. J., & Griffin, W. S. (1996), Neurobiologyof Aging, 17(5), 761-766).

Although the research to date may support a link between AD and IL-1, itis unclear what role IL-1 plays. IL-1 modulates actions that maycontribute to AD pathology, including the synthesis and processing ofthe Aβ precursor protein and the activity of acetylcholinesterase (Mrak,R. E. & Griffin, W. S. (2001), Neurobiology of Aging, 22(6), 903-908).Chronic IL-1 expression has been associated with demyelination (Ferrari,C. C., Depino, A. M., Prada, F., Muraro, N., Camptbell, S., Podhajcer,O., Pitossi, F. J. (2004), American Journal of Pathology, 165(5),1827-1837), breakdown of the blood-brain barrier, and neutrophilrecruitment (Ferrari et al, supra; Shaftel, S. S., Carlson, T. J.,Olschowka, J. A., Kyrkanides, S., Matousek, S. B., & O'Banion, M. K.(2007a), Journal of Neuroscience, 27(35), 9301-9309). IL-1 alsoactivates microglia, which in turn produce pro-inflammatory cytokinessuch as IL-1, IL-6, and tumor necrosis factor alpha (TNFa). Neuronalinsults, such as the accumulation of Aβ, may therefore induce aself-propagating cycle of cytokine activation in which levels of IL-1constantly rise, leading to neuronal damage and further plaquedeposition (Griffin, W. S. T., Sheng, J. G., Royston, M. C., Gentelman,S. M., McKenzie, J. E., Graham, D. I., Mrak, R. E. (1998), BrainPathology, 8, 65-72).

On the other hand, some have argued that IL-1 plays a beneficial role inAD. Sustained IL-1β overexpression for 4 weeks reduces amyloid plaqueexpression in swAPP-PS1 mice, a mouse model of Alzheimer's-likepathology that uses the Swedish pedigree mutation in the amyloidprecursor protein and the high Alzheimer's risk polymorphism inpresenilin-1 (Shaftel, S. S., Kyrkanides, S., Olschowka, J. A., Miller,J. H., Johnson, R. E., & O'Banion, M. K. (2007b), Journal of ClinicalInvestigation, 117(6), 1595-1604). In the AD brain, microglia expressingIL-1 surround amyloid plaque deposits, suggesting an attempt atphagocytic removal of the plaques (Griffin, W. S., Stanley, L. C., Ling,C., White, L., MacLeod, V., Perrot, L. J., Araoz, C. (1989), Proceedingsof the National Academy of Sciences of the United States of America, 86,7611-7615). In early stages of the disease, microglial activation doesseem to delay the progression of AD-like pathology (El Khoury, J., &Luster, A. D. (2008), Trends in Pharmacological Sciences, 29, 626-632;Simard, A. R., Soulet, D., Gowing, G., Julien, J., & Rivest, S. (2006),Neuron, 49, 489-502). However, amyloid plaque burden eventuallyincreases, despite continued microglial activation. One explanation isthat the microglia become defective and lose their Aβ-clearingeffectiveness. The expression of microglial Aβ receptors andAβ-degrading enzymes start to decrease around 8 months of age inswAPP-PS1 mice, resulting in reduced Aβ uptake and clearance (Hickman,S. E., Allison, E. K., & El Khoury, J. (2008), Neurobiology of Disease,28, 8354-8360). The microglia, however, maintain production of IL-1β andTNFα.

Hippocampally-mediated memory processes may be impaired by theoverexpression of IL-1 (Moore, A. H., Wu, M., Shaftel, S., Graham, K.A., & O'Banion, M. K. (2009), Neuroscience, 164, 1484-1495; Tanaka, S.,Ide, M., Shibutani, T., Ohtaki, H., Numazawa, S., Shioda, S., & Yoshida,T. (2006), Journal of Neuroscience Research, 83, 557-566; Depino, A. M.,Alonso, M., Ferrari, C., del Ray, A., Anthony, D., Besedovsky, H.,Pitossi, F. (2004), Hippocampus, 14, 526-535).

Although prior research has explored the use of anti-inflammatory drugsas a therapeutic strategy against AD, randomized clinical trials havefailed to consistently support the therapeutic effectiveness of NSAIDsagainst AD (Scharf, S., Mander, A., Ugoni, A., Vajda, F., &Christophidis, N. (1999), Neurology, 53(1), 197-201; Aisen, P. S.,Schafer, K. A., Grundman, M., Pfeiffer, E., Sano, M., Davis, K. L.,Thal, L. J. (2003), Journal of the American Medical Association,289(21), 2819-2826).

To date, there have been no studies examining the effects of chronic,systemic IL-1 inhibition on AD-like behavior and pathology. The currentstudies, described herein, utilized a mouse IL-1 Trap (mlL-1 Trap), animmunoadhesin consisting of a forced IL-1 receptor 1 homodimer fused toa mouse Fc fragment. This trap binds IL-1 at a high affinity, preventingit from binding to its endogenous receptor, and therefore serves as anantagonist of IL-1 signaling. After five months of subcutaneousadministration of the mlL-1 Trap in the swAPP-PS1 mutant mouse model ofAlzheimer's-like pathology, the effect of IL-1 inhibition on spatialmemory, open field activity, amyloid plaque burden, microglialactivation, and overall inflammation was examined.

Definitions

By the term “blocker”, “inhibitor”, or “antagonist” is meant a substancethat retards or prevents a chemical or physiological reaction orresponse. Common blockers or inhibitors include but are not limited toantisense molecules, antibodies, antagonists and their derivatives. Morespecifically, an example of an IL-1 blocker or inhibitor is an IL-1antagonist including, but not limited to, an antibody (human orhumanized), or an antigen binding portion thereof, to IL-1 alpha and/orIL-1 beta, a soluble IL-1 receptor that blocks or inhibits the activityof either IL-1 alpha or IL-1 beta or both, or an IL-1 trap as describedherein, which binds and inhibits IL-1 activity. The relevant IL-1 trapsthat may be used in the methods of the invention include any of theamino acid sequences noted in SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16,18, 20, 22, 24, 26 and 28.

By the term “therapeutically effective dose” is meant a dose thatproduces the desired effect for which it is administered. The exact dosewill depend on the purpose of the treatment, and will be ascertainableby one skilled in the art using known techniques (see, for example,Lloyd (1999) The Art, Science and Technology of PharmaceuticalCompounding).

By the term “substantially identical” is meant a protein sequence havingat least 95% identity to an amino acid sequence selected from the groupconsisting of the amino acid sequences SEQ ID NOs: 2, 4, 6, 8, 10, 12,14, 16, 18, 20, 22, 24, 26 and 28, and capable of binding IL-1 andinhibiting the biological activity of IL-1.

The term “identity” or “homology” is construed to mean the percentage ofamino acid residues in the candidate sequence that are identical withthe residue of a corresponding sequence to which it is compared, afteraligning the sequences and introducing gaps, if necessary to achieve themaximum percent identity for the entire sequence, and not consideringany conservative substitutions as part of the sequence identity. NeitherN- or C-terminal extensions nor insertions will be construed as reducingidentity or homology. Methods and computer programs for the alignmentare well known in the art. Sequence identity may be measured usingsequence analysis software (e.g., Sequence Analysis Software Package,Genetics Computer Group, University of Wisconsin Biotechnology Center,1710 University Ave., Madison, Wis. 53705). This software matchessimilar sequences by assigning degrees of homology to varioussubstitutions, deletions, and other modifications.

The term “treating” (or “treat” or “treatment”) refers to processesinvolving a slowing, interrupting, inhibiting, arresting, controlling,stopping, reducing, ameliorating, or reversing the progression,duration, or severity of an existing symptom, disorder, condition, ordisease, but does not necessarily involve a total elimination of alldisease-related symptoms, conditions, or disorders through use of theIL-1 trap as described herein. Furthermore, “treating”, “treatment” or“treat” refers to an approach for obtaining beneficial or desiredresults including clinical results, which include, but are not limitedto, one or more of the following: inhibiting, delaying or preventing theonset of, or the progression of, a disease associated with beta amyloidactivity, or characterized by aberrant deposition of beta amyloid in asubject, such as in Alzheimer's disease; or inhibiting, preventing, orameliorating at least one symptom associated with a disease associatedwith beta amyloid activity, or characterized by aberrant deposition ofbeta amyloid in a subject, such as in Alzheimer's disease, wherein thesymptoms include, but are not limited to, cognitive impairment, memoryloss, depression, anxiety, dementia, irritability, confusion, moodswings, aggressive and/or apathetic behavior. “Treatment” or “treating”,as used herein, also refers to increasing the quality of life of thosesuffering from the disease, decreasing the dose of other medicationsrequired to treat the disease and/or prolonging survival of patients.

“Delaying the onset of” Alzheimer's disease or a symptom thereof meansto defer, hinder, slow, retard, stabilize, and/or postpone developmentof the disease, or a symptom associated with, or resulting from thedisease. This delay can be of varying lengths of time, depending on thehistory of the disease and/or individual being treated. As is evident toone skilled in the art, a sufficient or significant delay can, ineffect, encompass prevention, in that the individual does not developthe disease. A method that “delays” development of Alzheimer's diseaseis a method that reduces probability of disease development in a giventime frame and/or reduces extent of the disease in a given time frame,when compared to not using the method. Such comparisons are typicallybased on clinical studies, using a statistically significant number ofsubjects.

“Development” of Alzheimer's disease means the onset and/or progressionof Alzheimer's disease within an individual. Alzheimer's diseasedevelopment can be detectable using standard clinical techniques.However, development also refers to disease progression that may beinitially undetectable. For purposes of this invention, progressionrefers to the biological course of the disease state, in this case, asdetermined by a standard neurological examination, patient interview, ormay be determined by more specialized testing. A variety of thesediagnostic tests include, but are not limited to, neuroimaging,detecting alterations of levels of specific proteins in the serum orcerebrospinal fluid (e.g., amyloid peptides and Tau), computerizedtomography (CT), positron emission tomography (PET), and magneticresonance imaging (MRI). “Development” includes occurrence, recurrence,and onset. As used herein “onset” or “occurrence” of Alzheimer's diseaseincludes initial onset and and/or recurrence.

The term “Alzheimer's Disease” or “AD” generally refers to a clinicalentity that typically presents with a characteristic progressiveamnestic disorder with subsequent appearance of other cognitive,behavioral and neuropsychiatric changes that impair social function andactivities of daily living. The initial presentation can be atypical,with non-amnestic focal cortical cognitive symptoms. Disease onsetand/or progression can now be assessed through the use of validated anddisease-specific biomarkers. Laboratory and neuroimaging biomarkers arehighly correlated with neuropathological lesions of AD. These biomarkerscan be divided into pathophysiological and topographical markers.Pathophysiological markers correspond to the two etiologicaldegenerative processes that characterize Alzheimer's pathology: theamyloidosis path to neuritic plaques and the tauopathy path toneurofibrillary tangles. They include CSF measurements of concentrationsof amyloid beta, total tau, and phosphotau, amyloid PET scanning withPittsburgh compound B or other radioligands (florbetaben, ¹⁸F-AV-45,etc.). Topographical markers are used to assess the less specific anddownstream brain changes that correlate with the regional distributionof AD pathology and include medial temporal lobe atrophy (as measured byMRI) and reduced glucose metabolism in temporo-parietal regions onfluorodeoxyglucose PET. These MRI and PET markers have been shown topredict the development of AD dementia in mild cognitive impairment(MCI) cohorts and to correlate with disease severity. Patients withclinical AD suffer from moderate to severe cognitive and memoryimpairments that meet the diagnostic criteria of AD and impact work andrelationships (including, potentially activities of daily living) andthese symptoms are usually accompanied by positive findings on abiomarker tests as described above.

“Prodromal Alzheimer's disease”, also referred to as “predementia stageof AD” refers to the early symptomatic predementia phase of AD inwhich 1) clinical symptoms including episodic memory loss of thehippocampal type are present, but not sufficiently severe to affectinstrumental activities of daily living and do not warrant a diagnosisof dementia; and in which 2) biomarker evidence from CSF or imaging issupportive of the presence of AD pathological changes.

“Preclinical Alzheimer's disease”, which includes both “asymptomaticat-risk state for AD” and “presymptomatic AD” refer to the longasymptomatic stage between the earliest pathogenic events/brain lesionsof AD and the first appearance of specific cognitive changes. The“asymptomatic at-risk” state for AD is identified in vivo by evidence ofamyloidosis in the brain (with retention of specific PET amyloidtracers) or in the CSF (with changes in amyloid beta, tau, andphosphotau concentrations). “Presymptomatic AD” applies to individualswho will develop AD and this can only be ascertained in families thatare affected by rare autosomal dominant monogenic mutations (monogenicAD).

“Cerebral amyloid angiopathy” (CAA), also known as congophilicangiopathy is a form of angiopathy in which amyloid deposits form in thewalls of the blood vessels of the central nervous system. The termcongophilic is used because the presence of the abnormal aggregations ofamyloid can be demonstrated by microscopic examination of brain tissueafter application of a special stain called Congo red. The amyloidmaterial is only found in the brain and as such the disease is notrelated to other forms of amyloidosis. CAA has been identified asoccurring either sporadically (generally in elderly populations) or infamilial forms such as Flemish, Iowa, and Dutch types. Sporadic forms ofCAA have been further characterized into two types based on depositionof amyloid β-protein (Aβ) in cortical capillaries. In all cases, it isdefined by the deposition of Aβ in the leptomeningal and cerebral vesselwalls. Amyloid deposition predisposes these blood vessels to failure,increasing the risk of a hemorrhagic stroke. Since this can be caused bythe same amyloid protein that is associated with Alzheimer's dementia,such brain hemorrhages are more common in people who suffer fromAlzheimer's, however they can also occur in those who have no history ofdementia. The hemorrhage within the brain is usually confined to aparticular lobe and this is slightly different compared to brainhemorrhages that occur as a consequence of high blood pressure(hypertension), a more common cause of a hemorrhagic stroke (or cerebralhemorrhage).

IL-1- Specific Antagonists

The invention provides IL-1 antagonists for the treatment of diseasescharacterized by aberrant deposition of beta amyloid in a subject, suchas in patients suffering from Alzheimer's disease (AD). In certainembodiments the IL-1 antagonists may include an antibody (or an antigenbinding fragment thereof) specific for IL-1 alpha or IL-1 beta, or asoluble receptor that blocks or inhibits the activity of IL-1 alpha orIL-1 beta or both. In certain embodiments, the IL-1 antagonist may beanakinra or canakinumab. In certain embodiments, the IL-1 antagonist maybe an IL-1-specific fusion protein antagonist (sometimes referred to asan “IL-1 trap”), which is useful for treating such conditions. IL-1traps are multimers of fusion proteins containing IL-1 receptorcomponents and a multimerizing component capable of interacting with themultimerizing component present in another fusion protein to form ahigher order structure, such as a dimer. Cytokine traps include twodistinct receptor components that bind a single cytokine, resulting inthe generation of antagonists with dramatically increased affinity overthat offered by single component reagents. In fact, the cytokine trapsthat are described herein are among the most potent cytokine blockersever described. Briefly, the cytokine traps called IL-1 traps arecomprised of the extracellular domain of human IL-1 R Type I (IL-1RI) orType II (IL-1RII) followed by the extracellular domain of human IL-1Receptor Accessory protein (IL-1RAcP), followed by a multimerizingcomponent. In one embodiment, the multimerizing component is animmunoglobulin-derived domain, such as, for example, the Fc region ofhuman IgG, including part of the hinge region, the CH2 and CH3 domains.An immunoglobulin-derived domain may be selected from any of the majorclasses of immunoglobulins, including IgA, IgD, IgE, IgG and IgM, andany subclass or isotype, e.g. IgG1, IgG2, IgG3 and IgG4; IgA-1 andIgA-2. For a more detailed description of the IL-1 traps, seeWO00/18932, U.S. Pat. No. 6,927,044; U.S. Pat. No. 7,459,426, whichpublications are herein specifically incorporated by reference in theirentirety. Preferred IL-1-specific fusion proteins have the amino acidsequence shown in SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22,24, 26 and 28, or a substantially identical protein at least 95%identity to a sequence of SEQ ID NO:4, 6, 8, 10, 12, 14, 16, 18, 20, 22,24, 26, or 28, and capable of binding and inhibiting IL-1.

In certain embodiments, the IL-1 antagonist comprises an antibodyfragment capable of binding IL-1a, IL-β, IL-1R1 and/or IL-1RAcp, or afragment thereof. The preferred embodiment would be an antagonist ofIL-1 R. One embodiment of an IL-1 antagonist comprises one or moreantibody fragments, for example, single chain Fv (scFv), is described inU.S. Pat. No. 6,472,179, which publication is herein specificallyincorporated by reference in its entirety. In all of the IL-1 antagonistembodiments comprising one or more antibody-derived components specificfor IL-1 or an IL-1 receptor, the components may be arranged in avariety of configurations, e.g., a IL-1 receptorcomponent(s)-scFv(s)-multimerizing component; IL-1 receptorcomponent(s)-multimerizing component-scFv(s); scFv(s)-IL-1 receptorcomponent(s)-multimerizing component, ScFv-ScFv-Fc, etc., so long as themolecule or multimer is capable of inhibiting the biological activity ofIL-1.

Anti-IL-1 Human Antibodies and Antibody Fragments

In another embodiment of the IL-1 antagonist useful in the method of theinvention, examples of anti-IL-1 antibodies are disclosed in U.S. Pat.No. 4,935,343; U.S. Pat. No. 5,681,933; WO 95/01997; EP 0267611, U.S.Pat. No. 6,419,944; WO 02/16436 and WO 01/53353. The IL-1 antagonist ofthe invention may include an antibody or antibody fragment specific foran IL-1 ligand (e.g., IL-1α or IL-1β) and/or an IL-1 receptor (e.g.,IL-1R1 and/or IL-1RAcp). Antibody fragments include any fragment havingthe required target specificity, e.g. antibody fragments either producedby the modification of whole antibodies (e.g. enzymatic digestion), orthose synthesized de novo using recombinant DNA methodologies (scFv,single domain antibodies or dAbs, single variable domain antibodies) orthose identified using human phase display libraries (see, for example,McCafferty et al. (1990) Nature 348:552-554). Alternatively, antibodiescan be isolated from mice producing human or human-mouse chimericantibodies using standard immunization and antibody isolation methods,including but not limited to making hybridomas, or using B cellscreening technologies, such as SLAM. Immunoglobulin binding domainsalso include, but are not limited to, the variable regions of the heavy(V_(H)) or the light (V_(L)) chains of immunoglobulins. Or by immunizingpeople and isolating antigen positive B cells and cloning the cDNAsencoding the heavy and light chain and coexpressing them in a cell, suchas CHO.

The term “antibody” as used herein refers to a polypeptide comprising aframework region from an immunoglobulin gene or fragments thereof thatspecifically binds and recognizes an antigen. The recognizedimmunoglobulin genes include the kappa, lambda, alpha, gamma, delta,epsilon, and mu constant regions, as well as the myriad immunoglobulinvariable region genes. Light chains are classified as either kappa orlambda. Heavy chains are classified as gamma, mu, alpha, delta, orepsilon, which in turn define the immunoglobulin classes, IgG, IgM, IgA,IgD, and IgE, respectively. Within each IgG class, there are differentisotypes (eg. IgG₁, IgG₂, IgG₃, IgG₄). Typically, the antigen-bindingregion of an antibody will be the most critical in determiningspecificity and affinity of binding.

An exemplary immunoglobulin (antibody) structural unit comprises atetramer. Each tetramer is composed of two identical pairs ofpolypeptide chains, each pair having one light chain (about 25 kD) andone heavy chain (about 50-70 kD). The N-terminus of each chain defines avariable region of about 100-110 or more amino acids primarilyresponsible for antigen recognition. The terms “variable light chain”(V_(L)) and variable heavy chain (V_(H)) refer to these light and heavychains respectively.

Antibodies exist as intact immunoglobulins, or as a number ofwell-characterized fragments produced by digestion with variouspeptidases. For example, pepsin digests an antibody below the disulfidelinkages in the hinge region to produce F(ab)′₂, a dimer of Fab whichitself is a light chain joined to V_(H)-C_(H)1 by a disulfide bond. TheF(ab)′₂ may be reduced under mild conditions to break the disulfidelinkage in the hinge region, thereby converting the F(ab)′₂ dimer intoan Fab′ monomer. The Fab′ monomer is essentially Fab with part of thehinge region. While various antibody fragments are defined in terms ofthe digestion of an intact antibody, one of skill will appreciate thatsuch fragments may be synthesized de novo either chemically or by usingrecombinant DNA methodology.

Methods for preparing antibodies are known to the art. See, for example,Kohler & Milstein (1975) Nature 256:495-497; Harlow & Lane (1988)Antibodies: a Laboratory Manual, Cold Spring Harbor Lab., Cold SpringHarbor, N.Y.). The genes encoding the heavy and light chains of anantibody of interest can be cloned from a cell, e.g., the genes encodinga monoclonal antibody can be cloned from a hybridoma and used to producea recombinant monoclonal antibody. Monoclonal antibodies can behumanized using standard cloning of the CDR regions into a humanscaffold. Gene libraries encoding human heavy and light chains ofmonoclonal antibodies can also be made from hybridoma or plasma cells.Random combinations of the heavy and light chain gene products generatea large pool of antibodies with different antigenic specificity.Techniques for the production of single chain antibodies or recombinantantibodies (U.S. Pat. No. 4,946,778; U.S. Pat. No. 4,816,567) can beadapted to produce antibodies used in the fusion proteins and methods ofthe instant invention. Also, transgenic mice, or other organisms such asother mammals, may be used to express human, human-mouse chimeric, orhumanized antibodies. Alternatively, phage display technology can beused to identify human antibodies and heteromeric Fab fragments thatspecifically bind to selected antigens.

Antibody Screening and Selection

Screening and selection of preferred antibodies can be conducted by avariety of methods known to the art. Initial screening for the presenceof monoclonal antibodies specific to a target antigen may be conductedthrough the use of ELISA-based methods, for example. A secondary screenis preferably conducted to identify and select a desired monoclonalantibody for use in construction of the multi-specific fusion proteinsof the invention. Secondary screening may be conducted with any suitablemethod known to the art. One preferred method, termed “BiosensorModification-Assisted Profiling” (“BiaMAP”) is described in co-pendingU.S. Ser. No. 60/423,017 filed 1 Nov. 2002, herein specificallyincorporated by reference in its entirety. BiaMAP allows rapididentification of hybridoma clones producing monoclonal antibodies withdesired characteristics. More specifically, monoclonal antibodies aresorted into distinct epitope-related groups based on evaluation ofantibody:antigen interactions. Antibodies capable of blocking either aligand or a receptor may be identified by a cell based assay, such as aluciferase assay utilizing a luciferase gene under the control of anNFKB driven promoter. Stimulation of the IL-1 receptors by IL-1 ligandsleads to a signal through NFKB thus increasing luciferase levels in thecell. Blocking antibodies are identified as those antibodies thatblocked IL-1 induction of luciferase activity.

Treatment Population

The therapeutic methods of the invention are useful for treatingindividuals affected with a disease or condition characterized byaberrant deposition and/or activity of beta amyloid in a subject.Diseases or conditions for which the current IL-1 antagonists may beused include Alzheimer's disease, multi-infarct dementia, cognitiveimpairment and cerebral amyloid angiopathy (CAA). The stages ofAlzheimer's disease (AD) or cerebral amyloid angiopathy (CAA) for whichthe treatments may be effective include any of the following: prodromalAD or CAA, preclinical AD or CAA and clinical stage AD or CAA, asdescribed previously.

Therapeutic Administration and Formulations

The invention provides therapeutic compositions comprising the IL-1antagonist (IL-1 trap) of the present invention. The administration oftherapeutic compositions in accordance with the invention will beadministered via a suitable route including, but not limited to,intravenously, subcutaneously, intramuscularly, intrathecally,intracerebrally, intraventricularly, intranasally, with suitablecarriers, excipients, and other agents that are incorporated intoformulations to provide improved transfer, delivery, tolerance, and thelike. A multitude of appropriate formulations can be found in theformulary known to all pharmaceutical chemists: Remington'sPharmaceutical Sciences, Mack Publishing Company, Easton, Pa. Theseformulations include, for example, powders, pastes, ointments, jellies,waxes, oils, lipids, lipid (cationic or anionic) containing vesicles(such as LIPOFECTIN™), DNA conjugates, anhydrous absorption pastes,oil-in-water and water-in-oil emulsions, emulsions carbowax(polyethylene glycols of various molecular weights), semi-solid gels,and semi-solid mixtures containing carbowax. See also Powell et al.“Compendium of excipients for parenteral formulations” PDA (1998) JPharm Sci Technol 52:238-311.

The dose of the IL-1 antagonist, e.g. IL-1 trap, may vary depending uponthe age and the size of a subject to be administered, target disease,conditions, route of administration, and the like. When the antibody ofthe present invention is used for treating Alzheimer's disease, it isadvantageous to intravenously administer the antibody of the presentinvention normally at a single dose of about 1 to about 750 mg/kg bodyweight, more preferably about 5 to about 300, about 10 to about 200, orabout 50 to about 150 mg/kg body weight. Depending on the severity ofthe condition, the frequency and the duration of the treatment can beadjusted. In certain embodiments, the IL-1 trap of the invention can beadministered as an initial dose of at least about 0.1 mg to about 800mg, about 1 to about 500 mg, about 5 to about 300 mg, or about 10 toabout 200 mg, to about 100 mg, or to about 50 mg. In certainembodiments, the initial dose may be followed by administration of asecond or a plurality of subsequent doses of the antibody orantigen-binding fragment thereof in an amount that can be approximatelythe same or less than that of the initial dose, wherein the subsequentdoses are separated by at least 1 day to 3 days; at least one week, atleast 2 weeks; at least 3 weeks; at least 4 weeks; at least 5 weeks; atleast 6 weeks; at least 7 weeks; at least 8 weeks; at least 9 weeks; atleast 10 weeks; at least 12 weeks; or at least 14 weeks. The IL-1 trapof the invention may be administered to a subject using any of the abovedescribed dosing regimens throughout the life of the patient.

Various delivery systems are known and can be used to administer thepharmaceutical composition of the invention, e.g., encapsulation inliposomes, microparticles, microcapsules, recombinant cells capable ofexpressing the mutant viruses, receptor mediated endocytosis (see, e.g.,Wu et al. (1987) J. Biol. Chem. 262:4429-4432). Methods of introductioninclude, but are not limited to, intradermal, transdermal,intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal,epidural, intrathecal, intraventricular, and oral routes. Thecomposition may be administered by any convenient route, for example byinfusion or bolus injection, by absorption through epithelial ormucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa,etc.) and may be administered together with other biologically activeagents. Administration can be systemic or local.

The pharmaceutical composition can be also delivered in a vesicle, inparticular a liposome (see, for example, Langer (1990) Science249:1527-1533).

In certain situations, the pharmaceutical composition can be deliveredin a controlled release system. In one embodiment, a pump may be used.In another embodiment, polymeric materials can be used. In yet anotherembodiment, a controlled release system can be placed in proximity ofthe composition's target, thus requiring only a fraction of the systemicdose.

The injectable preparations may include dosage forms for intravenous,subcutaneous, intracutaneous and intramuscular injections, dripinfusions, etc. These injectable preparations may be prepared by methodspublicly known. For example, the injectable preparations may beprepared, e.g., by dissolving, suspending or emulsifying the antibody orits salt described above in a sterile aqueous medium or an oily mediumconventionally used for injections. As the aqueous medium forinjections, there are, for example, physiological saline, an isotonicsolution containing glucose and other auxiliary agents, etc., which maybe used in combination with an appropriate solubilizing agent such as analcohol (e.g., ethanol), a polyalcohol (e.g., propylene glycol,polyethylene glycol), a nonionic surfactant [e.g., polysorbate 80,HCO-50 (polyoxyethylene (50 mol) adduct of hydrogenated castor oil)],etc. As the oily medium, there are employed, e.g., sesame oil, soybeanoil, etc., which may be used in combination with a solubilizing agentsuch as benzyl benzoate, benzyl alcohol, etc. The injection thusprepared is preferably filled in an appropriate ampoule.

A pharmaceutical composition of the present invention can be deliveredsubcutaneously or intravenously with a standard needle and syringe. Inaddition, with respect to subcutaneous delivery, a pen delivery devicereadily has applications in delivering a pharmaceutical composition ofthe present invention. Such a pen delivery device can be reusable ordisposable. A reusable pen delivery device generally utilizes areplaceable cartridge that contains a pharmaceutical composition. Onceall of the pharmaceutical composition within the cartridge has beenadministered and the cartridge is empty, the empty cartridge can readilybe discarded and replaced with a new cartridge that contains thepharmaceutical composition. The pen delivery device can then be reused.In a disposable pen delivery device, there is no replaceable cartridge.Rather, the disposable pen delivery device comes prefilled with thepharmaceutical composition held in a reservoir within the device. Oncethe reservoir is emptied of the pharmaceutical composition, the entiredevice is discarded.

Numerous reusable pen and autoinjector delivery devices haveapplications in the subcutaneous delivery of a pharmaceuticalcomposition of the present invention. Examples include, but certainlyare not limited to AUTOPEN™ (Owen Mumford, Inc., Woodstock, UK),DISETRONIC™ pen (Disetronic Medical Systems, Burghdorf, Switzerland),HUMALOG MIX 75/25™ pen, HUMALOG™ pen, HUMALIN 70/30™ pen (Eli Lilly andCo., Indianapolis, Inn.), NOVOPEN™ I, II and III (Novo Nordisk,Copenhagen, Denmark), NOVOPEN JUNIOR™ (Novo Nordisk, Copenhagen,Denmark), BD™ pen (Becton Dickinson, Franklin Lakes, N.J.), OPTIPEN™,OPTIPEN PRO™, OPTIPEN STARLET™, and OPTICLIK™ (sanofi-aventis,Frankfurt, Germany), to name only a few. Examples of disposable pendelivery devices having applications in subcutaneous delivery of apharmaceutical composition of the present invention include, butcertainly are not limited to the SOLOSTAR™ pen (sanofi-aventis), theFLEXPEN™ (Novo Nordisk), and the KWIKPEN™ (Eli Lilly), the SURECLICK™Autoinjector (Amgen, Thousands Oaks, Calif.), the PENLET™ (Haselmeier,Stuttgart, Germany), the EPIPEN (Dey, L.P.) and the HUMIRA™ Pen (AbbottLabs, Abbott Park, Ill.), to name only a few.

Advantageously, the pharmaceutical compositions for oral or parenteraluse described above are prepared into dosage forms in a unit dose suitedto fit a dose of the active ingredients. Such dosage forms in a unitdose include, for example, tablets, pills, capsules, injections(ampoules), suppositories, etc. The amount of the aforesaid antibodycontained is generally about 5 to about 750 mg per dosage form in a unitdose; especially in the form of injection, it is preferred that theaforesaid antibody is contained in about 5 to about 100 mg and in about10 to about 250 mg for the other dosage forms.

Combination Therapies

In numerous embodiments, the IL-1 antagonists of the present inventionmay be administered in combination with one or more additional compoundsor therapies. Combination therapy may be simultaneous or sequential. TheIL-1 antagonists of the invention may be combined with other IL-1antagonists, such as antibodies specific for IL-1 alpha or IL-1 beta (orantigen binding fragments thereof), a soluble IL-1 receptor, or otherdifferent IL-1 traps. The IL-1 traps of the invention may also be becombined with, for example, ARICEPT® (donepezil HCl), EXELON®(rivastigmine tartrate), RAZADYNE® (galantamine HBr), steroids, anakinra(KINARET®, Amgen) or canakinumab. The IL-1 traps of the invention mayalso be combined with anti-IL-18 drugs, such as for example, IL-18BP ora derivative, an IL-18 Trap, anti-IL-18, anti-IL-18R1, oranti-IL-18Racp. Other co-therapies include aspirin or other NSAIDs,steroids such as prednisolone, other inflammatory inhibitors such asinhibitors of caspase-1, p38, IKK1/2, CTLA-4lg, anti-IL-6 or anti-IL6Ra,an antibody specific for tau, an antibody specific for beta amyloid(such as solanezumab, gantenerumab, or bapineuzumab) or a microtubulestabilizers (such as epothilone B).

Administration Regimens

According to certain embodiments of the present invention, multipledoses of the IL-1 trap may be administered to a subject over a definedtime course. The methods according to this aspect of the inventioncomprise sequentially administering to a subject multiple doses of theIL-1 trap of the invention. As used herein, “sequentially administering”means that each dose of the IL-1 trap is administered to the subject ata different point in time, e.g., on different days separated by apredetermined interval (e.g., hours, days, weeks or months). The presentinvention includes methods, which comprise sequentially administering tothe patient a single initial dose of the IL-1 trap, followed by one ormore secondary doses of the IL-1 trap, and optionally followed by one ormore tertiary doses of the IL-1 trap.

The terms “initial dose,” “secondary doses,” and “tertiary doses,” referto the temporal sequence of administration of an IL-1 trap of theinvention. Thus, the “initial dose” is the dose which is administered atthe beginning of the treatment regimen (also referred to as the“baseline dose”); the “secondary doses” are the doses which areadministered after the initial dose; and the “tertiary doses” are thedoses which are administered after the secondary doses. The initial,secondary, and tertiary doses may all contain the same amount of theIL-1 trap, but generally may differ from one another in terms offrequency of administration. In certain embodiments, however, the amountof the IL-1 trap contained in the initial, secondary and/or tertiarydoses varies from one another (e.g., adjusted up or down as appropriate)during the course of treatment. In certain embodiments, two or more(e.g., 2, 3, 4, or 5) doses are administered at the beginning of thetreatment regimen as “loading doses” followed by subsequent doses thatare administered on a less frequent basis (e.g., “maintenance doses”).

In one exemplary embodiment of the present invention, each secondaryand/or tertiary dose is administered % to 26 (e.g., %, 1, 1%, 2, 2%, 3,3%, 4, 4%, 5, 5%, 6, 6%, 7, 7%, 8, 8%, 9, 9%, 10, 10%, 11, 11%, 12, 12%,13, 13%, 14, 14%, 15, 15%, 16, 16%, 17, 17%, 18, 18%, 19, 19%, 20, 20%,21, 21%, 22, 22%, 23, 23%, 24, 24%, 25, 25%, 26, 26%, or more) weeksafter the immediately preceding dose. The phrase “the immediatelypreceding dose,” as used herein, means, in a sequence of multipleadministrations, the dose of the IL-1 trap, which is administered to apatient prior to the administration of the very next dose in thesequence with no intervening doses.

The methods according to this aspect of the invention may compriseadministering to a patient any number of secondary and/or tertiary dosesof the IL-1 trap. For example, in certain embodiments, only a singlesecondary dose is administered to the patient. In other embodiments, twoor more (e.g., 2, 3, 4, 5, 6, 7, 8, or more) secondary doses areadministered to the patient. Likewise, in certain embodiments, only asingle tertiary dose is administered to the patient. In otherembodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8, or more) tertiarydoses are administered to the patient.

In embodiments involving multiple secondary doses, each secondary dosemay be administered at the same frequency as the other secondary doses.For example, each secondary dose may be administered to the patient 1 to2 weeks after the immediately preceding dose. Similarly, in embodimentsinvolving multiple tertiary doses, each tertiary dose may beadministered at the same frequency as the other tertiary doses. Forexample, each tertiary dose may be administered to the patient 2 to 4weeks after the immediately preceding dose. Alternatively, the frequencyat which the secondary and/or tertiary doses are administered to apatient can vary over the course of the treatment regimen. The frequencyof administration may also be adjusted during the course of treatment bya physician depending on the needs of the individual patient followingclinical examination.

Pharmaceutical Compositions

The present invention also provides pharmaceutical compositions. Suchcompositions comprise a therapeutically effective amount of an activeagent, and a pharmaceutically acceptable carrier. The term“pharmaceutically acceptable” means approved by a regulatory agency ofthe Federal or a state government or listed in the U.S. Pharmacopeia orother generally recognized pharmacopeia for use in animals, and moreparticularly, in humans. The term “carrier” refers to a diluent,adjuvant, excipient, or vehicle with which the therapeutic isadministered. Such pharmaceutical carriers can be sterile liquids, suchas water and oils, including those of petroleum, animal, vegetable orsynthetic origin, such as peanut oil, soybean oil, mineral oil, sesameoil and the like. Suitable pharmaceutical excipients include starch,glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silicagel, sodium stearate, glycerol monostearate, talc, sodium chloride,dried skim milk, glycerol, propylene, glycol, water, ethanol and thelike. The composition, if desired, can also contain minor amounts ofwetting or emulsifying agents, or pH buffering agents. Thesecompositions can take the form of solutions, suspensions, emulsion,tablets, pills, capsules, powders, sustained-release formulations andthe like. The composition can be formulated as a suppository, withtraditional binders and carriers such as triglycerides. Oral formulationcan include standard carriers such as pharmaceutical grades of mannitol,lactose, starch, magnesium stearate, sodium saccharine, cellulose,magnesium carbonate, etc. Examples of suitable pharmaceutical carriersare described in “Remington's Pharmaceutical Sciences” by E. W. Martin.

In one embodiment, the composition is formulated in accordance withroutine procedures as a pharmaceutical composition adapted forintravenous administration to human beings. Where necessary, thecomposition may also include a solubilizing agent and a local anestheticsuch as lidocaine to ease pain at the site of the injection. Where thecomposition is to be administered by infusion, it can be dispensed withan infusion bottle containing sterile pharmaceutical grade water orsaline. Where the composition is administered by injection, an ampouleof sterile water for injection or saline can be provided so that theingredients may be mixed prior to administration.

The active agents of the invention can be formulated as neutral or saltforms. Pharmaceutically acceptable salts include those formed with freeamino groups such as those derived from hydrochloric, phosphoric,acetic, oxalic, tartaric acids, etc., and those formed with freecarboxyl groups such as those derived from sodium, potassium, ammonium,calcium, ferric hydroxides, isopropylamine, triethylamine, 2-ethylaminoethanol, histidine, procaine, etc.

The amount of the active agent of the invention which will be effectivein the treatment of Alzheimer's disease can be determined by standardclinical techniques based on the present description. In addition, invitro assays may optionally be employed to help identify optimal dosageranges. The precise dose to be employed in the formulation will alsodepend on the route of administration, and the seriousness of thecondition, and should be decided according to the judgment of thepractitioner and each subject's circumstances. However, suitable dosageranges for intravenous administration are generally about 20 microgramsto 2 grams of active compound per kilogram body weight. Suitable dosageranges for intra-nasal administration are generally about 0.01 pg/kgbody weight to 1 mg/kg body weight. Effective doses may be extrapolatedfrom dose-response curves derived from in vitro or animal model testsystems.

For systemic administration, a therapeutically effective dose can beestimated initially from in vitro assays. For example, a dose can beformulated in animal models to achieve a circulating concentration rangethat includes the IC₅₀ as determined in cell culture. Such informationcan be used to more accurately determine useful doses in humans. Initialdosages can also be estimated from in vivo data, e.g., animal models,using techniques that are well known in the art. One having ordinaryskill in the art could readily optimize administration to humans basedon animal data.

Dosage amount and interval may be adjusted individually to provideplasma levels of the compounds that are sufficient to maintaintherapeutic effect. In cases of local administration or selectiveuptake, the effective local concentration of the compounds may not berelated to plasma concentration. One having skill in the art will beable to optimize therapeutically effective local dosages without undueexperimentation.

The amount of compound administered will, of course, be dependent on thesubject being treated, on the subject's weight, the severity of theaffliction, the manner of administration, and the judgment of theprescribing physician. The therapy may be repeated intermittently whilesymptoms are detectable or even when they are not detectable. Thetherapy may be provided alone or in combination with other drugs.

Kits

The invention also provides an article of manufacturing comprisingpackaging material and a pharmaceutical agent contained within thepackaging material, wherein the pharmaceutical agent comprises at leastone IL-1-specific fusion protein of the invention and wherein thepackaging material comprises a label or package insert which indicatesthat the IL-1-specific fusion protein can be used for treating a diseasecharacterized by aberrant deposition of beta amyloid, such asAlzheimer's disease.

Other features of the invention will become apparent in the course ofthe following descriptions of exemplary embodiments which are given forillustration of the invention and are not intended to be limitingthereof.

EXAMPLES

The following example is put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the methods and compositions of the invention, and are notintended to limit the scope of what the inventors regard as theirinvention. Efforts have been made to ensure accuracy with respect tonumbers used (e.g., amounts, temperature, etc.) but some experimentalerrors and deviations should be accounted for. Unless indicatedotherwise, parts are parts by weight, molecular weight is averagemolecular weight, temperature is in degrees Centigrade, and pressure isat or near atmospheric.

Example 1 Effect of an IL-1 Antagonist (IL-1 Trap) in a Mouse Model ofAlzheimer's-Like Pathology Methods Animals and Injections:

Subjects were 40 male mice split into two cohorts of 20. Within eachcohort, ten mice were wild type (WT) animals and ten were tandemtransgenic (Tg) for both the Swedish pedigree mutation in the amyloidprecursor protein (SwAPP) and the high Alzheimer's risk polymorphism inpresenilin-1 (PS-1). Both the wild type controls and the transgenic micewere of the C57B1/6 background. The mice were housed in atemperature-stabilized facility on a 12:12 light:dark cycle (lights on07:00), with food and water available ad libitum.

Starting at approximately 8 months of age, animals were administeredbiweekly injections of mouse IL-1 Trap or control mFc subcutaneously.Five Tg animals and five WT animals of each cohort received mlL-1 Trapat a dose of 10 mg/kg. The rest of the mice received mouse Fc (mFc) atthe same dose and volume. Mice were weighed weekly in order to establishdosage and evaluate animal health. The injections continued for 5months. Three mice died before completing the behavioral testing phase(one WT animal receiving mFc, one TG animal receiving mFc and one TGanimal receiving mlL-1 Trap.)

Behavioral testing:

Morris Water Maze: During the fifth month of injections, the MorrisWater Maze Test was conducted to evaluate spatial learning and memory. Apool measuring 105 cm in diameter and 35cm in depth was filled withwater made opaque with non-toxic white paint. The pool was then dividedconceptually into four quadrants, one of which contained an escapeplatform hidden 2.5 cm beneath the surface. Animals were placed in adifferent, pseudo-randomly selected quadrant at the start of each trial,and the latency to escape the maze onto the hidden platform wasmeasured. There were two trial blocks per day for five days, eachconsisting of three one-minute trials. Between trials, animals were heldwith a towel for approximately 30 seconds before being placed back inthe water. After each block, they were placed in a holding cage linedwith towels until dry and then returned to their home cages. The testingwas conducted at the same time each day with a 3-hour interval betweenthe first and second daily blocks. Any animal not locating the platformwithin one minute was assigned a latency of 60 seconds and guided to theplatform by hand before being removed from the water. Normal animalswere expected to demonstrate a decrease in latency to escape acrosstrials in the water maze, indicating acquisition of the location of theplatform over time.

One hour after the final block of water maze acquisition testing,animals were returned to the maze for a 30 second probe trial with theplatform removed to test their retention of the platform's location.Retention was assessed via a measure of the amount of time animals spentin the quadrant that formerly housed the platform, termed the “goalquadrant.” The number of times the animals swam over the spot where theplatform was located was also counted. Animals with normal retention areexpected to spend more time in the “goal quadrant” than in otherquadrants and to make more “platform crosses”, thus demonstratingretention of the spatial location of the escape platform.

For each block, the animals' median latency to escape the maze acrossthe three trials was recorded and these median latencies were used forstatistical analysis. In addition, animals' swim speeds were estimatedby dividing latency to escape by the number of maze quadrants crossedper trial to calculate mean quadrant crossing time. The median quadrantcrossing time for each trial block was used as a covariate in thestatistical analysis to account for potential differences in motorspeed.

Open Field:

During the fifth month, the open field test was conducted in order toexplore some of the non-cognitive behavioral symptoms ofAlzheimer's-like disease and whether IL-1 inhibition has an effect onthose symptoms. Specifically, open field can be used to measure generallocomotor and exploratory activity (Walsh, R. N. & Cummins, R. A.(1976), Psychological Bulletin, 83(3), 482-504). Animals were placedinto a white, box-like apparatus measuring 48 x 48×24 cm. The insidefloor was divided into nine grids, each measuring 16×16 cm. Animals wereplaced in the center grid and allowed to freely explore the apparatusfor a six minute trial. The number of total grid crossings was recordedas a measure of general locomotor and exploratory behavior.

Tissue Collection and Processing:

After 5 months of treatment, the animals were sacrificed and brains wereprepared for histological analysis. All animals were overdosed with apentobarbital-based euthanasia solution and perfusion fixed. Coldheparinized isotonic (0.9%) saline was run through the body toexsanguinate the animal and animals were then perfused with 4%paraformaldehyde first in acetate, and then borate buffer (100 ml each).Upon completion of the perfusion, brains were removed and placed in 30%buffered sucrose for 3-7 days. The fixed brains were sectioned coronallyat 50 μm and stored in cryoprotectant (Watson, R. E., Wiegand, S. J.,Clough, R. W., & Hoffman, G. E. (1986), Peptides, 7(1), 155-159) at −20°C. until they were stained.

Histology:

Brain tissue from the animals was used for histological analysis.Sections were stained in a 1:12 series with Congo Red to detect thepresence of amyloid plaques and with cresyl violet for Nissl bodies inseparate sections for visualization of all cells. Immunostaining wasconducted to visualize the microglial marker lba-1 (Millipore rabbitpolyclonal anti-lba-1, 1:500). For the second cohort of animals, adouble-stain was conducted for both plaques and microglia in order toevaluate parameters of microglia at various distances from plaques.

Image Analysis:

Amyloid plaque burden was analyzed by contrast analysis using the ImageJimage analysis software program (NIH). A minimum of 2 bilateral sectionsof hippocampus in a 1:12 series were selected and images were capturedin 10× using the PictureFrame program. In each picture, the red color ofthe stain was isolated and the background faded using Adobe Photoshop toachieve contrast. Identical processing parameters were used for allsections. The entire image was then converted to black and white andimported to Image J. For each image, a set percentage of the backgroundwas removed from all images and the image was converted to binary. Thepercentage of area stained, number of separate plaques, and average sizeper plaque were recorded for each animal.

The level of overall inflammation was assessed by subjective ratings. Anexperienced histologist blind to the experimental conditions examinedcresyl violet-stained hippocampal sections at a magnification of 40×.Each animal was given a rating of “none,” “mild,” “moderate,” or“marked,” based on the presence of inflammatory cell profiles(microglia, immune cells), with “none” signifying no inflammatory cellsand “severe” being the highest amount of inflammation observed. Thepresence of microglial-invested deposits was also noted. For each animalshowing plaque-like deposits, 5 random deposits were selected from thehippocampus and the surrounding microglia-like cells were counted. Themedian number of microglial profiles per plaque was recorded for eachanimal. Finally, double stains for congo red-positive plaques andlba-1-immunoreactive microglia were used to measure the sizes ofmicroglia contacting plaques (contact), within 20 microns of a plaque(adjacent), or greater than 30 microns from a plaque (distant).

Statistical Analysis

For water maze acquisition, the median latency to escape to platform wasrecorded for each animal for each trial block. A three-way mixedFactorial analysis of variance (ANOVA) was conducted using treatment andgenotype as the between groups independent variables and block as arepeated measure. For retention, the proportion of time spent in thegoal quadrant was recorded for each animal along with the number ofplatform crosses. Two-way ANOVAs (genotype×treatment) were conductedwith each of these as dependent variables.

To evaluate locomotor activity and exploratory behavior, the number ofgrid crossings in the open field test was counted for each animal. Atwo-way factorial ANOVA was conducted using treatment and genotype asthe between groups independent variables.

Two-way ANOVAs (genotype×treatment) were conducted for each of thefollowing measures: amyloid plaque total area, plaque number,hippocampal volume, microglial size, and average plaque size. For thesemeasures, values for individual hemispheres were averaged to calculate amean for each animal. Finally, regression analyses were conducted toassess the predictive relationship between memory performance and plaquepathology for each treatment group (mFc and mlL-1 Trap).

A non-parametric log-linear analysis for qualitative variables wasconducted to analyze the overall inflammation ratings. An independentgroups t-test was conducted to compare number of microglia-like cellssurrounding hippocampal deposits in mFc-treated transgenic mice versusmlL-1-treated trap transgenic mice.

Data are presented as mean across all animals within atreatment/genotype group and standard error of the mean (SEM).Statistical significance was set at an alpha level of 0.05.

Results General Health:

Mice were weighed and evaluated weekly to assess their general health.Out of the original 40 animals, 37 survived to sacrifice. In the firstcohort, one transgenic animal in the mFc-treated group died before thebehavioral testing stage. From the second cohort, one transgenic animalin the mlL-1 Trap group died before the behavioral testing stage and onewild type animal in the mlL-1 Trap group died after completing somebehavioral testing. At the final weighing before perfusion, thetransgenic mice weighed about 27% less than the wild type mice(F_((1,33))=55.46, p<0.001). Although the transgenic mice weighed less,they were actually closer to the normal body weight for adult male mice,and appeared grossly healthy. The mlL-1 Trap had no negative impact onbody weights (F_((1,33))=0.00, p=0.994) and the mice all seemed healthythroughout the study.

Water Maze:

As expected, a three-way mixed Factorial ANOVA (genotype×treatment×trialblock) showed that the wild type animals performed significantly betteroverall than the swAPP/PS-1 double transgenic animals in water mazeacquisition (F_((1,33))=29.71, p<0.001). The mlL-1 Trap had nosignificant overall effect on water maze performance (F_((1,33))=2.76,p=0.11), but there was a significant interaction between the two factors(F_((1,33))=4.61, p=0.039; FIG. 1), such that the trap selectivelyimproved performance in the transgenic animals. An analysis of cohorteffects revealed a significant cohort by genotype interaction, such thatthe transgenic animals of the second cohort performed worse overall thanthose in the first cohort. However, there was no cohort by treatmenteffect and no cohort by genotype by treatment effect, indicating thatthe trap affected the mice similarly across both cohorts.

We also assessed whether differences in swim speed could be influencingthese results. Slower swim speeds have been reported in transgenic ADmice (Ying, T., Xu, Y., Scearce-Levie, K., Ptacek, L. J., & Fu, Y. H.(2010), Neurogenetics, 11(1), 41-52), which can interfere with ananimal's latency to swim to the platform. A three-way mixed FactorialANOVA (genotype×treatment×trial block) revealed no significantdifferences in quadrant crossing time across genotype (F_((1,33))=0.61,p=0.442) or treatment (F_((1,33))=1.13, p=0.296), and no genotype bytreatment interaction (F_((1,33))=0.47, p=0.50). In addition, we did notsee any significant general locomotor differences in the open field test(see Open Field section), suggesting that motor differences cannotaccount for the acquisition results observed.

On the retention portion of the water maze, wild type animals spentsignificantly more time in the goal quadrant (F_((1,33))=18.83, p<0.001)and made significantly more platform crosses (F_((1,33))=8.86, p=0.005)than transgenic animals, confirming the memory impairment in thetransgenics. There was no significant effect of IL-1 Trap treatment onproportion of time spent in the goal quadrant (F_((1,33))=0.313, p=0.58)and there was no interaction between the factors (F_((1,33))=0.04,p=0.84) (FIG. 1b ). There was also no significant effect of treatment onnumber of platform crosses (F_((1,33))=2.29, p=0.14) and no interactionbetween treatment and genotype (F_((1,33))=.1.63, p=0.21) (FIG. 1c ).

Open Field

A two-way ANOVA (genotype×treatment) was conducted to examinedifferences in locomotor activity and exploratory behavior. Overall,there was no difference in total number of grid crossings between wildtype and transgenic animals (F_((1,33))=0.699, p=0.409), and nodifference between animals treated with mFc and those treated with mll-1Trap (F_((1,33))=0.18, p=0.673). There was no interaction betweengenotype and treatment (F_((1,33))=0.03, p=0.873). These resultsindicate that transgenic mice did not differ in overall level oflocomotor activity compared to wild type mice and that level of activitywas not affected by treatment with mlL-1 trap.

Plaque Burden Analysis

As expected, the plaque burden contrast analysis revealed thatswAPP/PS-1 transgenic mice had significantly more plaques(F(1,13)=34.21, p<0.001), more total area covered by plaques(F(1,13)=31.04, p<0.001), and a higher average plaque size(F(1,13)=7.43, p=0.02) than the wild type animals. Indeed, no Congored-positive plaques were observed in any wild type animal (data notshown). There was no significant difference between animals given mlL-1Trap and mFc on any of these measures, and no interactions betweengenotype and treatment (FIG. 3).

We also examined the relationship between plaque burden and spatiallearning acquisition on block 5 (halfway through learning) and block 10(final performance block) of water maze acquisition. For animals givenmFc, there was a significant positive correlation between total plaquearea and latency to escape on block 5 (r(6)=0.90, p=0.002), such thatanimals with more plaque coverage took longer to find the platformduring the learning trial. The correlation remained significant whenwild type animals, which had no plaques, were excluded from analysis(r(2)=0.96, p=0.04). A regression analysis could not be conducted toregress plaques against learning in the wild type animals, because noplaques were detected in any of these animals. The mlL-1 Trap animalshad a smaller, non-significant correlation between plaque burden andlearning performance (r(7)=0.46, p=0.21), which disappeared completelywhen wild type animals were excluded from analysis (r(3)=0.004, p=0.99),suggesting that mlL-1 Trap eliminated the association between plaqueburden and cognitive performance in the swAPP/PS-1 transgenic mice. Byblock 10, when animals had reached peak performance, the amount ofplaque coverage was no longer significantly correlated with water mazeperformance for either treatment group. Sample sizes were too small todetermine whether the differences in correlations between the twogenotypes were statistically significant.

Inflammation

A log-linear analysis of overall subjective inflammation ratings did notreveal a significant difference between mFc and mlL-1 Trap-treatedanimals (G²(4)=5.22, p=0.26) (Table 1). An independent groups t-test wasconducted on the measure of the overall number of microglia-like cellssurrounding the plaque-like deposits in the hippocampi of transgenicanimals, and there was no significant difference in microglial profilecounts between the Fc and mlL-1 Trap-treated animals. (t₍₇₎=−0.30,p=0.78) (FIG. 4a ). However, an analysis of microglial profile sizeshowed a statistically significant change in microglial size with IL-1Trap treatment interacting with distance from plaques (FIG. 4b ,interaction F(2,12)=16.127, p=0.0004). Specifically, while microglialsizes were larger for microglia contacting plaques in all animals, thedifferences in average size for plaques contacting microglia weresignificantly greater in IL-1 Trap-treated animals. To better illustratethis effect, the difference between the mean size of microglia inunaffected tissue versus plaque-containing tissue for each animal wascalculated. This difference in microglial size per animal showed astatistically significant effect of IL-1 Trap on enhancement inmicroglial size upon plaque contact (FIG. 4c , t(6)=4.864, p=0.0028).

In addition, as shown in FIG. 5, this model of disease produces nosignificant effect on overall hippocampal volume (F(1,13)=0.236,p=0.635), nor does IL-1 Trap impact hippocampal volume in either thewild type or transgenic mice (treatment F(1,13)=0.800, p=0.387;interaction F(1,13)=0.015, p-0.903).

TABLE 1 Distribution of Overall Inflammation Ratings None Mild ModerateMarked WT mFc 40% 60% 0% 0% mIL-1 Trap 20% 20% 60% 0% TG mFc 0% 25% 25%50% mIL-1 Trap 20% 50% 20% 20%

Discussion

Chronic neuroinflammation, which includes elevated IL-1 expression, is aprominent feature of Alzheimer's disease, which may contribute to theneurodegeneration and associated cognitive dysfunction observed inpatients with Alzheimer's Disease. In this study, systemicadministration of the mlL-1 Trap was used to inhibit interleukin-1signaling for 5 months after the onset of disease in order to observeits effects on both behavior and Alzheimer's-like brain pathology. Thestudy was done to determine if IL-1 inhibition would improve performanceon measures of learning and memory while reducing amyloid plaque burden.Although the mlL-1 Trap did improve water maze performance in transgenicmice, it did not significantly alter amyloid plaque burden. It did,however, increase the size of microglia contacting amyloid plaques whiledecreasing the size of microglia overall in the brain, suggesting thatalthough it didn't alter the amount of beta-amyloid deposited in thetransgenic brains, it may have altered the nature of the immune responsetriggered by the plaques. In addition, mlL-1 Trap completely eliminatedthe significant positive relationship between plaque burden andcognitive impairments, showing that although the plaques were present,they were no longer associated with cognitive impairments in theanimals.

Water Maze

Our finding that the mlL-1 Trap selectively improved water mazeperformance for transgenic animals shows that IL-1 inhibition can slowthe cognitive decline resulting from overexpression of mutantPresenilin-1 and the Swedish APP mutation. In the present study, IL-1inhibition improved acquisition (e.g. learning) while having no effecton retention (e.g. memory), showing that the treated mice may have usedcompensatory strategies to assist with their performance of the task.

Open Field

The open field test is used to evaluate locomotor and exploratorybehaviors in rodents. Results in swAPP-PS1 transgenic mice showed nosignificant difference in locomotor activity compared to wild type micein a six-minute open field test. The lack of significant differencesbetween groups on both open field and on water maze swim speed suggestthat differences in learning and memory performance cannot be attributedto alterations in motor or exploratory behavior.

Amyloid Plaques

In addition to the behavioral testing as a measure of Alzheimer's likepathology, a Congo Red stain was performed for the detection of amyloidplaques. We hypothesized that chronic IL-1 inhibition would result inreduced amyloid plaque pathology. However, the mlL-1 Trap did notsignificantly decrease hippocampal plaque burden in the transgenicanimals.

The water maze data taken together with the plaque analysis provides aninteresting look at the nature of AD-like pathology in the swAPP-PS1transgenic mice. If, as our data indicate, the Trap did improve spatialmemory in these mice without reducing the amount of amyloid plaques inthe brain, it would imply that factors other than β-amyloid depositionare contributing to their cognitive deficits. One explanation is thatthe mlL-1 Trap may be improving memory by inhibiting the immune responseto the plaques without reducing the plaques themselves.

To further evaluate the role of amyloid plaques, we looked at therelationship between plaque deposits and water maze performance fortransgenic animals treated with mFc and for animals treated with mlL-1Trap. In this study, transgenic AD animals given mFc showed a verystrong significant correlation (r=0.96) between plaque burden and watermaze acquisition performance during spatial memory learning (block 5),such that animals with more amyloid deposition took longer to find theplatform. However, animals given mlL-1 Trap showed no correlation(r=0.004) between plaque burden and the same water maze measure. Thefact that these results were obtained with such small sample sizes isespecially promising, although further studies with larger sample sizeare warranted. If the mlL-1 Trap, which suppresses part of theneuroinflammatory response, actually eliminates the relationship betweenamyloid plaques and memory, it would support the hypothesis that it isthe immune response to the plaques, rather than the plaques themselves,that are causing the memory impairments.

Inflammation

In order to probe the possibility that mlL-1 Trap changed the immuneresponse to the plaques, a subjective analysis of the overall level ofinflammation around the plaques in the hippocampi of the transgenicanimals was conducted. The initial analysis involved the assignment ofan overall subjective rating of peri-plaque inflammation by anexperienced histologist blind to animal treatment. This analysis did notreveal any significant overall differences in inflammatory cellinvestment around the plaques. In support of this finding, aquantitative count of one specific inflammatory cell type, the residentbrain macrophages (microglia), revealed no difference in overallmicroglial count per plaque. However, a statistical trend was observedtoward a significant difference in microglial size between transgenicanimals treated with IL-1 Trap versus mFc control. The differences thatwere observed included a shift toward smaller microglia at rest, butlarger microglia contacting plaques. Although more research will benecessary to fully interpret this trend, one possibility is thatmicroglia are less activated basally in IL-1 Trap treated animals, butare more phagocytic when confronted with pathological deposits. While wedo not have direct evidence that IL-1 Trap changed the activationsubtype of microglia, this pattern of sizes may indicate that more M2than M1 microglia were present in the brains of IL-1 Trap-treatedanimals. M2 microglia tend to be less activated overall, but have morepotential for phagocytosis.

It is possible that the inhibition of interleukin 1 in the present studyis affecting some of the detrimental microglial processes, withoutimproving their plaque-clearing ability. Further morphometric andneurochemical analysis will help determine the full extent of changes inmicroglial activation and morphology.

Summary

The current study showed a potentially protective role of systemic mlL-1Trap treatment in the swAPP/PS-1 double transgenic model of Alzheimer'sDisease. In particular, the current study provides support for thegrowing hypothesis that amyloid plaques do not, in themselves, underliethe hallmark cognitive impairments of Alzheimer's Disease. Indeed, itsupports the idea that components of the inflammatory cascade, perhapstriggered in part by the presence of the plaques, are major pathogeniccontributors. In addition, our data provide initial proof of concept forthe potential use of IL-1 inhibition to treat the cognitive impairmentsof Alzheimer's Disease. Finally, our data show that even largebiological inhibitors of IL-1, such as the IL-1 traps described herein,given after disease onset, could provide significant benefit to patientssuffering from the profound cognitive impairments characterizing thisdevastating disease.

1. A method for treating, or delaying the onset, or the progression of adisease characterized in part by beta amyloid expression, activity, ordeposition in a subject in need thereof, or for ameliorating at leastone symptom associated with the disease, the method comprisingadministering to the subject a therapeutically effective amount of anIL-1 antagonist as a first therapeutic agent, wherein the IL-1antagonist is selected from the group consisting of an antibody specificfor IL-1 alpha or IL-1 beta, or an antigen-binding fragment thereof, asoluble IL-1 receptor, and an IL-1 trap, wherein the IL-1 trap is afusion protein comprising an IL-1 binding portion of the extracellulardomain of IL-1RAcP, an IL-1 binding portion of the extracellular domainof IL-1R1, and a multimerizing component.
 2. A method for treating, ordelaying the onset, or the progression of a disease characterized inpart by beta amyloid expression, activity, or deposition in a subject inneed thereof, or for ameliorating at least one symptom associated withthe disease, the method comprising administering to the subject atherapeutically effective amount of an IL-1 antagonist as a firsttherapeutic agent, wherein the IL-1 antagonist is an IL-1 trap, whereinthe IL-1 trap is a fusion protein comprising an IL-1 binding portion ofthe extracellular domain of IL-1RAcP, an IL-1 binding portion of theextracellular domain of IL-1R1, and a multimerizing component.
 3. Themethod of either claim 1 or 2, wherein the subject is a human.
 4. Themethod of claim 1, wherein the disease is selected from the groupconsisting of Alzheimer's disease (AD), Down's syndrome, multi-infarctdementia, cognitive impairment and cerebral amyloid angiopathy (CAA). 5.The method of claim 4, wherein the Alzheimer's disease is clinical,pre-clinical or prodromal Alzheimer's disease.
 6. The method of claim 4,wherein the cerebral amyloid angiopathy is clinical or pre-clinicalcerebral amyloid angiopathy.
 7. The method of claim 1, wherein the IL-1antagonist is an IL-1 trap comprising a fusion protein having an aminoacid sequence selected from the group consisting of SEQ ID NOs: 2, 4, 6,8, 10, 12, 14, 16, 18, 20, 22, 24, 26, and 28, or a substantiallyidentical sequence having at least 95% identity to the sequence selectedfrom the group consisting of SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 18,20, 22, 24, 26, and 28 and capable of binding and inhibiting IL-1. 8.The method of claim 7, wherein the IL-1 trap is a fusion proteincomprising the amino acid sequence of SEQ ID NO:10 or SEQ ID NO:
 28. 9.The method of claim 1, wherein administration is subcutaneous,intramuscular, intranasal, intraarterial, intravenous, intrathecal,intraventricular, intracerebral, topical, transdermal administration ororal.
 10. The method of claim 1, wherein a therapeutically effectiveamount is between about 1 mg/kg to about 750 mg/kg.
 11. The method ofclaim 10, wherein a therapeutically effective amount is between about 10mg/kg to about 500 mg/kg.
 12. The method of claim 11, wherein atherapeutically effective amount is between about 50 mg/kg to about 150mg/kg.
 13. The method of claim 1, further comprising administering to asubject in need thereof a therapeutically effective amount of one ormore other therapeutic agents, wherein the disease or at least onesymptom associated with the disease is lessened in severity or duration,or wherein the onset or progression of the disease or at least onesymptom associated with the disease is delayed.
 14. The method of claim1, wherein the at least one symptom associated with the disease isselected from the group consisting of memory loss, depression, anxiety,inattention, dementia, irritability, confusion, mood swings, andaggressive and/or apathetic behavior.
 15. The method of claim 13,wherein administration of the other therapeutic agent is subcutaneous,intramuscular, intranasal, intraarterial, intravenous, intrathecal,intraventricular, intracerebral, topical, transdermal administration ororal.
 16. The method of claim 13, wherein the other therapeutic agent isan acetylcholinesterase inhibitor or a glutamate pathway modifier. 17.The method of claim 16, wherein the acetylcholinesterase inhibitor isselected from the group consisting of ARICEPT® (donepezil HCl), EXELON®(rivastigmine tartrate), and RAZADYNE® (galantamine HBr).
 18. The methodof claim 16, wherein the glutamate pathway modifier is Namenda(memantine).
 19. The method of claim 13, wherein the other therapeuticagent is selected from the group consisting of a different IL-1antagonist, an anti-inflammatory agent, an antibody specific for tau, anantibody specific for beta amyloid and a microtubule stabilizer.
 20. Themethod of claim 19, wherein the different IL-1 antagonist is selectedfrom the group consisting of an IL-1 alpha or IL-1 beta antibody, asoluble IL-1 receptor, a different IL-1 trap, anakinra and canakinumab.21. The method of claim 19, wherein the anti-inflammatory agent isaspirin or a different NSAID.
 22. The method of claim 19, wherein theantibody specific for beta amyloid is selected from the group consistingof solanezumab, gantenerumab, and bapineuzumab.
 23. The method of claim19, wherein the microtubule stabilizer is epothilone.
 24. A method ofimproving cognitive impairment in a mammal having beta amyloid depositsin brain tissue, the method comprising administering to the subject atherapeutically effective amount of an IL-1 antagonist as a firsttherapeutic agent, wherein the IL-1 antagonist is selected from thegroup consisting of an antibody specific for IL-1 alpha or IL-1 beta, oran antigen binding fragment thereof, a soluble IL-1 receptor, and anIL-1 fusion protein (IL-1 trap) comprising an IL-1 binding portion ofthe extracellular domain of IL-1RAcP, an IL-1 binding portion of theextracellular domain of IL-1R1, and a multimerizing component, whereinthe mammal demonstrates an improvement in cognitive function(s) withoutnecessarily exhibiting a concurrent change in the beta amyloid plaqueburden in the brain.
 25. A method of improving cognitive impairment in amammal having beta amyloid deposits in brain tissue, the methodcomprising administering to the subject a therapeutically effectiveamount of an IL-1 antagonist as a first therapeutic agent, wherein theIL-1 antagonist is an IL-1 fusion protein (IL-1 trap) comprising an IL-1binding portion of the extracellular domain of IL-1RAcP, an IL-1 bindingportion of the extracellular domain of IL-1R1, and a multimerizingcomponent, wherein the mammal demonstrates an improvement in cognitivefunction(s) without necessarily exhibiting a concurrent change in thebeta amyloid plaque burden in the brain.
 26. The method of claim 24,wherein the IL-1 antagonist is an IL-1 trap comprising a fusion proteinhaving an amino acid sequence selected from the group consisting of SEQID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, and 28, or asubstantially identical sequence having at least 95% identity to thesequence selected from the group consisting of SEQ ID NOs: 2, 4, 6, 8,10, 12,14, 16, 18, 20, 22, 24, 26, and 28 and capable of binding andinhibiting IL-1.
 27. The method of claim 25, wherein the IL-1 trap is afusion protein comprising the amino acid sequence of SEQ ID NO:10 or SEQID NO:
 28. 28. The method of claim 24, further comprising administeringto a subject in need thereof a therapeutically effective amount of oneor more other therapeutic agents, wherein the disease or at least onesymptom associated with the disease is lessened in severity or duration,or wherein the onset or progression of the disease or at least onesymptom associated with the disease is delayed.
 29. The method of claim28, wherein the at least one symptom associated with the disease isselected from the group consisting of memory loss, depression, anxiety,dementia, inattention, irritability, confusion, mood swings, andaggressive and/or apathetic behavior.
 30. The method of claim 28,wherein the administration of the other therapeutic agent issubcutaneous, intramuscular, intranasal, intraarterial, intravenous,intrathecal, intraventricular, intracerebral, topical, transdermaladministration or oral.
 31. The method of claim 28, wherein the othertherapeutic agent is an acetylcholinesterase inhibitor or a glutamatepathway modifier.
 32. The method of claim 31, wherein theacetylcholinesterase inhibitor is selected from the group consisting ofARICEPT® (donepezil HCl), EXELON® (rivastigmine tartrate), and RAZADYNE®(galantamine HBr).
 33. The method of claim 31, wherein the glutamatepathway modifier is Namenda (memantine).
 34. The method of claim 28,wherein the one or more other therapeutic agents are selected from thegroup consisting of a different IL-1 antagonist, an anti-inflammatoryagent, an antibody specific for tau, an antibody specific for betaamyloid and a microtubule stabilizer.
 35. The method of claim 34 whereinthe different IL-1 antagonist is selected from the group consisting ofan IL-1 alpha or IL-1 beta antibody, a soluble IL-1 receptor, adifferent IL-1 trap, anakinra and canakinumab.
 36. The method of claim34, wherein the anti-inflammatory agent is aspirin or a different NSAID.37. The method of claim 34, wherein the antibody specific for betaamyloid is selected from the group consisting of solanezumab,gantenerumab, and bapineuzumab.
 38. The method of claim 34, wherein themicrotubule stabilizer is epothilone.